×
| SKU | EK0525 |
|---|---|
| Size | 96wells/kit, with removable strips. |
| Reactivity | Human |
| Sample Type | cell culture supernates, serum and plasma(heparin, EDTA, citrate). |
| Sensitivity | <1pg/ml |
| Assay Range | 15.6pg/ml-1000pg/ml(cellculturesupernates),7.8pg/ml-500pg/ml(serum,plasma) |
Overview
| Product Name | Human TNF Alpha PicoKine™ ELISA Kit |
|---|---|
| SKU/Catalog Number | EK0525 |
| Storage & Handling | Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.) |
| Size | 96wells/kit, with removable strips. |
| Description | Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human TNF alpha. 96wells/kit, with removable strips. |
| Cite This Product | Human TNF Alpha PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0525) |
| Sample Type | cell culture supernates, serum and plasma(heparin, EDTA, citrate)..
Anticoagulant(s): heparin, EDTA or citrate
*The recommended anticoagulants are proven to not block the antibody binding sites on the target antigen. Please do not collect blood sample with other anticoagulants that are not specified above or contact us to check for feasibility. |
| Sensitivity | <1pg/ml |
| Assay Range | 15.6pg/ml-1000pg/ml(cellculturesupernates),7.8pg/ml-500pg/ml(serum,plasma) |
| Immunogen | Expression system for standard: E.coli, Immunogen sequence: V77-L233 |
| Reactivity | Human |
| Cross Reactivity | There is no detectable cross-reactivity with other relevant proteins. |
| Antibody Clonalities | Capture Antibody | Detection Antibody: monoclonal antibody from mouse|polyclonal antibody from goat |
Assay Details
Kit Components
| Catalog number | Description | Quantity |
| EK0525-CAP | 96-well plate precoated with anti-Human TNF antibody | 1 |
| EK0525-ST | lyophilized recombinant Human TNF standard | 10ng/tube |
| EK0525-DA | biotinylated anti-Human TNF antibody | 130ul |
| AR1103 | Avidin-Biotin-Peroxidase Complex(ABC) | 130ul |
| AR1106-1 | sample diluent buffer | 30ml |
| AR1106-2 | antibody diluent buffer | 12ml |
| AR1106-3 | ABC diluent buffer | 12ml |
| AR1104 | TMB color developing agent | 10ml |
| AR1105 | TMB stop solution | 10ml |
| PLA-SEA | Adhesive cover | 4 |
Materials Required But Not Provided
- Microplate reader in standard size.
- Automated plate washer.
- Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
- Clean tubes and Eppendorf tubes.
- Washing buffer (neutral PBS or TBS).
- Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g NaCl; 450μl of purified acetic acid or 700μl of concentrated hydrochloric acid to 1000ml H2
- Preparation of 0.01 M PBS: Add 8.5g sodium chloride, 1.4g Na O and adjust pH to 7.2-7.
Typical Data Obtained from Human TNF Alpha PicoKine™ ELISA Kit
(TMB reaction incubate at 37°C for 15-25min)
| Concentration(pg/ml) | 0 | 15.6 | 31.2 | 62.5 | 125 | 250 | 500 | 1000 |
| O.D. | 0.062 | 0.127 | 0.177 | 0.255 | 0.468 | 0.873 | 1.265 | 1.953 |
Intra/Inter Assay Precision
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean(pg/ml) | 16 | 82 | 201 | 14 | 75 | 186 |
| Standard deviation | 0.76 | 5.49 | 12.66 | 0.75 | 5.55 | 11.9 |
| CV(%) | 4.8% | 6.7% | 7.4% | 5.4% | 7.4% | 6.4% |
Reproducibility
Three samples with differing target protein concentrations were assayed using four different lots to measure the CV% lot to lot variance.
Reproducibility
To assay reproducibility, three samples with differing target protein concentrations were assayed using four different lots.
| Lots | Lot1 (pg/ml) | Lot2 (pg/ml) | Lot3 (pg/ml) | Lot4 (pg/ml) | Mean (pg/ml) | Standard Deviation | CV (%) |
|---|---|---|---|---|---|---|---|
| Sample 1 | 16 | 15 | 15 | 16 | 15 | 0.5 | 3.3% |
| Sample 2 | 82 | 91 | 75 | 89 | 84 | 6.29 | 7.4% |
| Sample 3 | 201 | 204 | 215 | 219 | 209 | 7.46 | 3.5% |
*The typical data is obtained from Boster's internal QC result and for reference only. It may differ from the lab test results of the end users. It is more important that the user's lab test results reflect the same linearity demonstrated in the typical data than achieving exactly the same O.D. values.
Target Info
Protein Target Info (Source: Uniprot.org)
| Uniprot Id | P01375 |
|---|---|
| Gene Name | TNF |
| Protein Name | Tumor necrosis factor |
| Alternative Names | Tumor necrosis factor;Cachectin;TNF-alpha;Tumor necrosis factor ligand superfamily member 2;TNF-a;Tumor necrosis factor, membrane form;N-terminal fragment;NTF;Intracellular domain 1;ICD1;Intracellular domain 2;ICD2;C-domain 1;C-domain 2;Tumor necrosis factor, soluble form;TNF;TNFA, TNFSF2; |
| Subcellular Localization | Cell membrane ; Single-pass type II membrane protein . |
| Molecular Weight | 25644 MW |
*if product is indicated to react with multiple species, protein info is based on the human gene.
Ontology
| Protein Function | Cytokine that binds to TNFRSF1A/TNFR1 and TNFRSF1B/TNFBR. It is mainly secreted by macrophages and can induce cell death of certain tumor cell lines. It is potent pyrogen causing fever by direct action or by stimulation of interleukin-1 secretion and is implicated in the induction of cachexia, Under certain conditions it can stimulate cell proliferation and induce cell differentiation. Impairs regulatory T-cells (Treg) function in individuals with rheumatoid arthritis via FOXP3 dephosphorylation. Upregulates the expression of protein phosphatase 1 (PP1), which dephosphorylates the key 'Ser-418' residue of FOXP3, thereby inactivating FOXP3 and rendering Treg cells functionally defective (PubMed:23396208). Key mediator of cell death in the anticancer action of BCG-stimulated neutrophils in combination with DIABLO/SMAC mimetic in the RT4v6 bladder cancer cell line (PubMed:22517918). . |
|---|---|
| Research Areas | Atherosclerosis, Cancer, Cardiovascular, Cytokines, Growth Factors, Growth Factors/Hormones, Immunology, Innate Immunity, Metabolism, Signal Transduction, Tnf Superfamily, Vascular Inflammation *You can search these to find other products in these research areas. |
| Background | Tumor necrosis factor-alpha(TNF-alpha, or TNF) is secreted by macrophages in response to inflammation, infection and cancer. Human Tumor Necrosis Factor(TNF) and Lymphotoxin(TNF-beta) are cytotoxic proteins which have similar biological activities and share 30% amino acid homology. TNF-alpha is produced by monocytes, which can stimulate endothelial cells to produce the multilineage growth factor granulocyte-macrophage colony-stimulating factor and extend the role of this immunoregulatory protein to the regulation of hematopoiesis in vitro. TNF is a soluble protein that causes damage to tumor cells but has no effect on normal cells. Human TNF has been purified to apparent homogeneity as a 17.3-kilodalton protein from HL-60 leukemia cells and has showed cytotoxic and cytostatic activities against various human tumor cell lines. The human TNF cDNA is 1585 base pairs in length and encodes a protein of 233 amino acids. The mature protein begins at residue 77, leaving a long leader sequence of 76 amino acids. TNF-alpha has been mapped to human chromosome 6. |
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Publications
Hypericin-photodynamic therapy induces human umbilical vein endothelial cell apoptosis
Expression of autophagy-related genes in cerebrospinal fluid of patients with tuberculous meningitis
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Oroxylin A inhibits colitis by inactivating NLRP3 inflammasome
Effect of different anesthetic methods on cellular immune functioning and the prognosis of patients with ovarian cancer undergoing oophorectomy
Prion protein is required for tumor necrosis factor ? (TNF?)-triggered nuclear factor ?B (NF-?B) signaling and cytokine production
hsdS, Belonging to the Type I Restriction-Modification System, Contributes to the Streptococcus suis Serotype 2 Survival Ability in Phagocytes
Wogonoside inhibits invasion and migration through suppressing TRAF2/4 expression in breast cancer
Heme Oxygenase-1, a Key Enzyme for the Cytoprotective Actions of Halophenols by Upregulating Nrf2 Expression via Activating Erk1/2 and PI3K/Akt in EA.hy926 Cells
Effect of Helix aspersa extract on TNF?, NF-?B and some tumor suppressor genes in breast cancer cell line Hs578T
Proinflammatory effects of S100A8/A9 via TLR4 and RAGE signaling pathways in BV-2 microglial cells
Effect of etomidate on the oxidative stress response and levels of inflammatory factors from ischemia-reperfusion injury after tibial fracture surgery
Imidazoline I2 receptor inhibitor idazoxan regulates the progression of hepatic fibrosis via Akt-Nrf2-Smad2/3 signaling pathway
Hypaphorine Attenuates Lipopolysaccharide-Induced Endothelial Inflammation via Regulation of TLR4 and PPAR-? Dependent on PI3K/Akt/mTOR Signal Pathway
Protective effect of geraniol inhibits inflammatory response, oxidative stress and apoptosis in traumatic injury of the spinal cord through modulation of NF-?B and p38 MAPK
An in vitro 3D bone metastasis model by using a human bone tissue culture and human sex-related cancer cells
Randomized, double-blind, placebo-controlled superiority trial of the Yiqigubiao pill for the treatment of patients with chronic obstructive pulmonary disease at a stable stage
miR-181a Induces Macrophage Polarized to M2 Phenotype and Promotes M2 Macrophage-mediated Tumor Cell Metastasis by Targeting KLF6 and C/EBP?
Evaluation of the effect of Vaccinium arctostaphylos L fruit extract on serum inflammatory biomarkers in adult hyperlipidemic patients: a randomized double-blind placebo-controlled clinical trial
Evaluation of optimized continuous venovenous hemodiafiltration therapy efficiency in severe burn patients with sepsis
LL202 protects against dextran sulfate sodium-induced experimental colitis in mice by inhibiting MAPK/AP-1 signaling
Pharmacological Inactivation of Src Family Kinases Inhibits LPS-Induced TNF-? Production in PBMC of Patients with Beh?et's Disease
NF-?B-Regulated miR-99a Modulates Endothelial Cell Inflammation.
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The Burkholderia cenocepacia OmpA-like protein BCAL2958: identification, characterization, and detection of anti-BCAL2958 antibodies in serum from B cepacia complex-infected Cystic Fibrosis patients
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Associations of Trauma Severity with Mean Platelet Volume and Levels of Systemic Inflammatory Markers (IL1?, IL6, TNF?, and CRP)
Salivary levels of interleukin-6 and tumor necrosis factor-? in patients undergoing hemodialysis
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Metabolic changes of H2S in smokers and patients of COPD which might involve in inflammation, oxidative stress and steroid sensitivity
Evaluation of adenosine deaminase (ADA) isoenzymes activity and tumor necrosis factor-? (TNF?) concentration in chronic heart failure
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Evaluation of the Inflammatory Response in Macrophages Stimulated with Exosomes Secreted by?Mycobacterium avium-Infected Macrophages
Apolipoprotein CIII regulates lipoprotein-associated phospholipase A2 expression via the MAPK and NF?B pathways
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NR4A1 is associated with chronic low-grade inflammation in patients with type 2 diabetes
RNF26 Temporally Regulates Virus-Triggered Type I Interferon Induction by Two Distinct Mechanisms
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Role of Pentoxifylline and Sparfloxacin in Prophylaxis of Spontaneous Bacterial Peritonitis in Cirrhotic Patients
TNF-? promotes gallbladder cancer cell growth and invasion through autocrine mechanisms
Evaluation of salivary tumor necrosis factor-alpha in patients with the chronic periodontitis: A case-control study
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TRIM38 inhibits TNF?- and IL-1??triggered NF-?B activation by mediating lysosome-dependent degradation of TAB2/3
GEC-derived SFRP5 Inhibits Wnt5a-Induced Macrophage Chemotaxis and Activation
Modulatory Effect of 1,25-Dihydroxyvitamin D3 on IL1 ?-Induced RANKL, OPG, TNF ?, and IL-6 Expression in Human Rheumatoid Synoviocyte MH7A
Effects of Pantoprazole on Systemic and Gastric Pro- and Anti-inflammatory Cytokines in Critically Ill Patients
Effect of n-3 and n-6 unsaturated fatty acids on prostate cancer (PC-3) and prostate epithelial (RWPE-1) cells in vitro
Cloning and expression analysis of a novel high-mobility group box 2 homologue from Lampetra japonica
Correlation between the microinflammatory state and left ventricular structural and functional changes in maintenance haemodialysis patients
Does Quercetin Improve Cardiovascular Risk factors and Inflammatory Biomarkers in Women with Type 2 Diabetes: A Double-blind Randomized Controlled Clinical Trial
Dynamic changes in HMGB1 levels correlate with inflammatory responses during cardiopulmonary bypass
Drug Screening for Autophagy Inhibitors Based on the Dissociation of Beclin1-Bcl2 Complex Using BiFC Technique and Mechanism of Eugenol on Anti-Influenza A Virus Activity
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Identification and Characterization of the Lamprey High-Mobility Group Box 1 Gene
Glucose and Fatty Acid Metabolism in a 3 Tissue In-Vitro Model Challenged with Normo- and Hyperglycaemia
Malnutrition and inflammation in acute kidney injury due to earthquake-related crush syndrome
Malnutrition and inflammation in acute kidney injury due to earthquake-related crush syndrome
Wu GR, Mu TC, Gao ZX, Wang J, Sy MS, Li CY. J Biol Chem. 2017 Nov 17;292(46):18747-18759. doi: 10.1074/jbc.M117.787283. Epub 2017 Sep 12. Prion protein is required for tumor necrosis factor α (TNFα)-triggered nuclear factor κB (NF-κB) signaling and cytokine production
Xu B, Zhang P, Li W, Liu R, Tang J, Fan H. Front Microbiol. 2017 Aug 9;8:1524. doi: 10.3389/fmicb.2017.01524. eCollection 2017. hsdS, Belonging to the Type I Restriction-Modification System, Contributes to the Streptococcus suis Serotype 2 Survival Ability in Phagocytes
Koh Y, Park KS. J Exerc Rehabil. 2017 Aug 29;13(4):472-476. doi: 10.12965/jer.1735066.533. eCollection 2017 Aug. Responses of inflammatory cytokines following moderate intensity walking exercise in overweight or obese individuals
Huang N, Xu Y, Zhou H, Lin D, Zhang B, Zhang Y, Pan D, Tao L, Liu X, Shen X. Med Sci Monit. 2017 Oct 4;23:4760-4767. Essential Oil from Fructus Alpiniae Zerumbet Protects Human Umbilical Vein Endothelial Cells In Vitro from Injury Induced by High Glucose Levels by Suppressing Nuclear Transcription Factor-Kappa B Signaling
Sun HJ, Chen D, Wang PY, Wan MY, Zhang CX, Zhang ZX, Lin W, Zhang F. Oxid Med Cell Longev. 2017;2017:6905217. doi: 10.1155/2017/6905217. Epub 2017 Nov 19. Salusin-β Is Involved in Diabetes Mellitus-Induced Endothelial Dysfunction via Degradation of Peroxisome Proliferator-Activated Receptor Gamma
Liu B, Piao X, Guo L, Wang G, Sun W, Gao L, Zheng X, Fang Y. Evid Based Complement Alternat Med. 2017;2017:7323129. doi: 10.1155/2017/7323129. Epub 2017 Nov 20. A New Chinese Medicine Intestine Formula Greatly Improves the Effect of Aminosalicylate on Ulcerative Colitis
Wen Y, Gan H, Li Z, Sun X, Xiong Y, Xia Y Sci Rep. 2018 Apr 13;8(1):5941. doi: 10.1038/s41598-018-24397-w. Safety of Low-calcium Dialysate and its Effects on Coronary Artery Calcification in Patients Undergoing Maintenance Hemodialysis
Min M, Yan BX, Wang P, Landeck L, Chen JQ, Li W, Cai SQ, Zheng M, Man XY. PLoS One. 2017 Dec 22;12(12):e0190051. doi: 10.1371/journal.pone.0190051. eCollection 2017. Rottlerin as a therapeutic approach?in psoriasis: Evidence from in vitro and in vivo studies
Xuanfei L, Hao C, Zhujun Y, Yanming L, Jianping G. Oncotarget. 2017 Mar 28;8(13):21015-21030. doi: 10.18632/oncotarget.15472. Imidazoline I2 receptor inhibitor idazoxan regulates the progression of hepatic fibrosis via Akt-Nrf2-Smad2/3 signaling pathway
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Han XR, Wen X, Li YY, Fan SH, Zhang ZF, Li H, Sun XF, Geng GQ, Sun S, Huang SQ, Wu DM, Lu J, Zheng YL. Biosci Rep. 2017 Oct 24;37(5). pii: BSR20170915. doi: 10.1042/BSR20170915. Print 2017 Oct 31. Effect of different anesthetic methods on cellular immune functioning and the prognosis of patients with ovarian cancer undergoing oophorectomy
Feng XE, Liang TG, Gao J, Kong P, Ge R, Li QS,. Oxid Med Cell Longev. 2017;2017:7028478. doi: 10.1155/2017/7028478. Epub 2017 Jun 14. Heme Oxygenase-1, a Key Enzyme for the Cytoprotective Actions of Halophenols by Upregulating Nrf2 Expression via Activating Erk1/2 and PI3K/Akt in EA.hy926 Cells
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T T Liao1, Q Y Deng1, B J Wu1, S S Li1, X Li1, J Wu1, Y X Leng1,3, Y B Guo2 and N Huang1
Published 24 January 2017 • © 2017 IOP Publishing Ltd
Biomedical Materials, Volume 12, Number 1
Dose-dependent cytotoxicity evaluation of graphite nanoparticles for diamond-like carbon film application on artificial joints
Isil Cakir1 / Eser Kilic2 / Banu Demet Ozel Coskun3 / Nuri Cakir
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Soluble IL-1 decoy receptor is associated with gastric adenocarcinoma
Zoology & Entomology Department, Faculty of Science, Helwan University and
2Department of Endemic Medicine & Hepatology, Faculty of Medicine, Cairo
University, Egypt
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Zhengwang Cao, Yiliang Fang, Yonghui Lu, Dunxian Tan, Changhong Du, Yuming Li, Qinlong Ma, Junmei Yu, Mengyan Chen, Chao Zhou, Liping Pei, Lei Zhang, Haiying Ran, Mindi He, Zhengping Yu, hou Zhou
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www.ijcep.com /ISSN:1936-2625/IJCEP0033609
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Department of Orthopedics, The Second Hospital of Tangshan, Tangshan, Hebei 063000, P.R. China
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DOI: 10.3892/etm.2016.3850
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Customer Q&As
Q: The volumes of my samples are small, would it be possible to use 50µl/sample?
A: There are two solutions for insufficient sample volume.
1. Diluent the samples with the provided sample diluent buffer into 100ul.
2. Add 50ul of standard solution, when 50ul of sample will be added into a well.
1. Diluent the samples with the provided sample diluent buffer into 100ul.
2. Add 50ul of standard solution, when 50ul of sample will be added into a well.
Q: Will this kit work on tissue samples?
A: For tissue homogenates, endogenous biotin should be considered.
Endogenous biotin in tissue homogenates might introduce false positive signal.
Please run the test as described below to confirm if there is Endogenous biotin in the tissue lysate samples using the ELISA kit.
• Add tissue homogenates into the wells and then add ABC and TMB without adding any biotinylated detection antibody to see if any signal will be observed.
• If no signal is produced, then you can work on the tissue sample by using the kit.
• Add tissue homogenates into the wells and then add ABC and TMB without adding any biotinylated detection antibody to see if any signal will be observed.
• If no signal is produced, then you can work on the tissue sample by using the kit.
Q: Is it possible to dilute 10X TBS stock that is 250mM Tris Base/Tris-Hcl, 27 mM KCl, and 1.37M NaCl to be used as the wash buffer?
A: You can dilute the 10X TBS stock to 1XTBS and test the pH value of the 1xTBS.
The 1XTBS can be used if the pH value falls in the range of 7.2-7.6.
The 1XTBS can be used if the pH value falls in the range of 7.2-7.6.
Q: Would your kits work normall with samples from Wistar Rat?
A: Our kits would work normally with samples from Wistar Rat.
Q: How can I store the dissolved wash buffer for a longer period of time?
A: You can prepare a concentrated wash buffer (25X) which can be stored at 4°C for 1-3 months.
Q: Does EK0525, EK0392, EK0373 react with BSA?
A: EK0525, EK0392, EK0373 does not react with BSA.
Q: For EK0525, EK0392, EK0373, do these kits have binding for the forms of cytokine other than just the active form?
A: EK0525, EK0392, EK0373 are able to detect total forms of TNF-Alpha, IL-1Beta, and IFN-Gamma.
Q: Does the TMB color developing agent contain H2O2?
A: Yes, the TMB color developing agent contain H2O2.
Q: Does EK0525 work with whole cell lysates?
A: EK0525 is not suitable for working with whole cell lysates.
Q: Does this ELISA kit contain any product produced in humans or primates?
Keywords: component, ingredient
Keywords: component, ingredient
A: None of the components in our ELISA kits are produced in humans or primates.
Q: What is the volume of the recombinant protein control? What is its shelf life?
Keywords: expiration, storage, temperature, size
Keywords: expiration, storage, temperature, size
A: The volume of the control is around 100pg. If unopened, the shelf life of the control is the same as the whole kit - "Store at 4˚C for 6 months, at -20˚C for 12 months." The reconstituted control can be stored at -20˚C for 2 days.
Q: Will it be okay to run the experiment if I accidentally used the wrong buffer (e.g. sample diluent buffer) for antibody dilution instead of the antibody diluent buffer? Keywords: dilution, replace, substitute, sample buffer
A: Using the sample diluent buffer for antibody dilution instead of the antibody diluent buffer will negatively affect the test result to some extent. The values of both standard and sample might be lower than the normal values.
Q: The protocol says to use neutral PBS and provides a recipe. Could I use neutral PBS made with KCl or KH2PO4 (common constituents for most PBS recipes) instead?
Keywords: protocol, alternative buffer, applications, reagent recipe
A: Yes, it is ok to use common PBS recipes. We have tried many types of buffers with common constituents, and no significant difference was observed whatsoever.
Q: Why are my O.D. values different than your values on the datasheet?
Keyword: troubleshooting, reference
A: We detected the kit in our lab and got our values on the datasheet before delivery, but protein activity will decrease as time goes on, so you may get lower O.D. values than ours. However, it should still be in a reasonable range and the standards can be used to calculate sample values. Good linearity of curve is more important than the actual numerical value.
Q: On the ELISA kit datasheet it says "HRP substrate TMB was used to visualize HRP enzymatic reaction." What is HRP and which part of the kit contains HRP?
Keyword: kit components, reagents, protocol
A: It means Avidin-Biotin-Peroxidase Complex(AR1103) and you could find it in Kit Components.
Q: Do you offer any ELISA kits that can work with whole blood samples instead of plasma or urine?
Keyword: applications
A: We test serum and plasma routinely, and there is very little difference between serum, plasma and whole blood. The whole blood also contains proteins we need to test. The kit can be used to test whole blood in theory.
Q: Can I calculate the results of unknown samples using a computer program with a 4-parameter logistic function?
Keyword: data analysis
A: Yes, 4-parameter logistic function can be used to calculate sample data.
Q: Does silicic acid (formula SiOH4) affect the results of the ELISA assay?
Keyword: protocol, reagents
A: The acid may affect the binding of antigen to antibody, it is not recommended to use. That being said, it is unlikely to affect the reaction if the solution remains an overall neutral environment.
Q: Is there lot-to-lot variation of the ELISA kits? What are Boster's general services when I have questions about your kits?
Keyword: technical support, help, customer service
A: The NIBSC/WHO 1st International Standard is evaluated, we always test our kits before delivery, and customers can find the test result on the protocol. If the customer needs technical support from us to analyze their data, please contact us and we ask that you provide us the following information: the lot# and production date, and when did the customer detect the kit.
Q: The results of my standards do not look correct, what could be the problem?
Keyword: troubleshoot, protocol
A: Double check the protocol for plate washing. It should be examined on three aspects: 1) Did you make the washing buffer correctly? 2) How long did you let the buffer stay in wells? 3) What was the volume of the washing buffer added to each well? In addition, pay attention when adding sample to avoid contamination. And we suggest testing the standards again. Values of standards at low concentration are more affected than that at higher concentration, so customer can still get expected values at high concentration even if there is an error. If problems persist, please contact technical support with the catalog and lot number of your product.
Q: What is the well depth of the 96 well plates for the ELISA kits?
Keyword: well capacity, product size
A: The well depth is 300ul, and the max capacity is 350ul. The height of the well is 1.1 cm, and the internal size is 1 cm.
Q: The kit does not include wash solution, what should I do?
Keyword: wash buffer
A: Our Elisa kits do not come with wash solution, but we have included information about how to make washing buffer on the datasheet. Please refer to the "Material Required But Not Provided" section. We also offer washing buffer for sale separately (Phosphate Buffered Saline Powder SKU: AR0030).
Q: For your ELISA kits, are the capture and detection antibodies polyclonals or monoclonals?
Keyword: antibody clonality
A: This information can be found for each kit under the "Properties" section, and you can find the immunogen sequence information in the "Overview" section.
Q: Do your ELISA kits come with sealants or plate covers?
Keyword: storage, sequential use
A: The kit can be used within a month sequentially if it's opened and stored at 4 degree. There is no need to use sealants, the plate can be packaged with aluminum foil bag, and for other reagents keep bottle tightly closed.
Q: What is the concentration of Sodium Azide in your Elisa kits?
Keyword: preservative
A: Our Elisa kits contain 0.02% Sodium Azide. All of the components except TMB colour developing reagent and stop solution contains 0.02% Sodium azide as the preservative.
Q: Are there positive and negative controls available for my ELISA kit?
Keyword: positive control, negative control
A: We can provide a recombinant protein as a control. It costs $50 per control and takes 2 weeks to manufacture. We cannot provide a positive or negative control in serum.
Q: Why do I get positive results for my knockout (KO) model when used as a control?
A: The knockout (KO) model may contain truncated forms of the target protein which can be detected by the ELISA assay.
Q: How long do I soak my plate in the wash buffer?
A: The plate should soak in 0.3mL PBS or TBS wash buffer for at least 1-2 minutes in an automated wash station.
Q: Can I use Tween in my wash buffer?
A: While it is not recommended to use Tween in your wash buffer, small amounts (<0.1% concentration) may decrease background due to insufficient blocking.
Q: What plate type do I use to set up the microplate reader?
A: Our plates are made with the Corning costar plates similar to the DNA-BIND 96 -well plates.
Q: What should I use for negative control?
A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the normal level of this protein in my sample of interest?
A: We have reviewed literature and have gathered this information for most of our ELISA kits. You can find this information on the product page or contact us if you cannot find it. However this information is only suggestive and cannot replace pilot studies in determining the optimal sample dilution ratio.
Q: Is the plate separable? Can I use only part of the kit and save the rest for later? How many samples can I run with one kit?
A: Yes the plate is separable. There are 12 strips of 8 wells each, all removable from the plate. The amount of samples you can run depend on a few factors. In the most common ELISA set up, you will use two strips for standards, and 10 strips for samples using duplicates, which let you run up to 40 samples per kit. Contact us if you have questions regarding other situations.
Q: Does the kit contain sample preparation reagents? How do I prepare my samples?
A: Since different sample types require different reagents, we do not include them in the ELISA kit. However we do have each reagent mentioned in the file below available at very reasonable prices. Be sure to check them out. For sample preparation protocols please download the file below: https://www.bosterbio.com/media/pdf/Boster_Sample_Preparation_Protocols.pdf
Q: Can this ELISA kit work on brain tissue homogenate, cell culture supernatant, saliva, urine, serum, whole blood or any other sample type?
A: In theory the ELISA kit will work for all sample types if the target protein is present at a level that falls within the linear range of the ELISA kit detection range. We guarantee the kit to work on the sample types that we have tested. If you want dilution ratio suggestions on these sample types please contact our technical support. For sample types that we have not tested for, we suggest you run pilot experiments to decide the optimal sample dilution.
Q: Can this ELISA kit react with the pro-form of the target protein? Can this ELISA kit react with an isoform of the protein?
A: In general, unless otherwise specified, the ELISA kit is pan-specific, meaning that it will react with all different forms of the target protein if they share the majority of the target protein's sequence. The capture and detection antibodies are reactive to the entire sequence of the standard protein. You can find the sequence information of the standard protein in the "immunogen" section. For more information about the specificity of the kit for your particular experiment, please contact our techincal support.
Q: Can this ELISA kit react with human, mouse, rat or other species?
A: If the kit is reactive to another commonly used species (human, mouse, and/or rat), we would have listed it as a separate product. If you want to check cross-reactivity to a species that is not included in the 3 species listed above, please contact our technical support for more information. As a rule of thumb, if the sequences are 90%+ identical, there is a high chance of cross-reactivity for your species of interest.
Q: What are some alternative names that could be used to describe this product?
A: Some common names include but are not limited to human tnf alpha elisa kit, human tnf-alpha elisa kit, human tnf elisa kit, human tnf a elisa kit, human tnf-a elisa kit