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SKU:EK0525
Pack Size:96wells/kit, with removable strips.
Application:ELISA
Sensitivity:<1pg/ml
Sample Type:cell culture supernates, serum and plasma(heparin, EDTA, citrate).
Price: $250.00
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Overview

Product Name Human TNF Alpha PicoKine™ ELISA Kit
SKU/Catalog Number EK0525
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)
Validated Species Human
Application ELISA

*Our Boster Guarantee covers the use of this product in the above tested applications.

**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.

Immunogen Expression system for standard: E.coli,V77-L233
Cross Reactivity There is no detectable cross-reactivity with other relevant proteins.
Pack Size 96wells/kit, with removable strips.

Properties

Sensitivity <1pg/ml
*Sensitivity, or Lower Limit of Detection (LLD), is the minimum level of target protein the ELISA assay can detect. We measure 20 blank wells and if the O.D. value is 2 standard deviations higher than the blanks' average O.D. the sample can be deemed positive.
Assay Range 15.6pg/ml-1000pg/ml(cellculturesupernates),7.8pg/ml-500pg/ml(serum,plasma)
*This assay range is determined using common samples. For samples with low target protein concentrations, users can adjust the standard curve to extend the lower limit of assay range.
Sample Type cell culture supernates, serum and plasma(heparin, EDTA, citrate).

*The above listed samples are the ones valided with the assay. If you do not see your sample of interest listed, as long as there is enough level of target protein present in the sample, this Picokine™ ELISA kit should detect it.
**For protocol and tips regarding preparing your sample of interest, please check our ELISA sample preparation guide.
Capture Antibody monoclonal antibody from mouse
Detection Antibody polyclonal antibody from goat
Natural Levels Some research articles suggesting the natural levels of TNF are listed below:
Storage Store at 4˚C for 6 months, at -20˚C for 12 months. Avoid multiple freeze-thaw cycles(Shipped with wet ice.)

Kit Components

Catalog numberDescriptionQuantity
EK0525-CAP96-well plate precoated with anti-Human TNF antibody1
EK0525-STlyophilized recombinant Human TNF standard10ng/tubex2
EK0525-DAbiotinylated anti-Human TNF antibody130ul(dilution 1:100)
AR1103Avidin-Biotin-Peroxidase Complex(ABC)130ul(dilution 1:100)
AR1106-1sample diluent buffer30ml
AR1106-2antibody diluent buffer12ml
AR1106-3ABC diluent buffer12ml
AR1104TMB color developing agent10ml
AR1105TMB stop solution10ml
PLA-SEAAdhesive cover4
AR0030-EPBS washing bufferPowder for 1000ml

Washing buffer Preparation: Disolve AR0030-E to 1000ml distilled water and adjust pH to 7.2~7.6. Finally, adjust the total volume to 1L.

*Additional components can be purchased. If you need extra of the above components please order them together to avoid additional shipping charges.

See Prices For Extra Components

Material Required But Not Provided

  • 1. Microplate reader in standard size.
  • 2. Automated plate washer.
  • 3. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection.
  • 4. Clean tubes and Eppendorf tubes.
*Reagents needed for sample collection and preparation are not included in the kit.

Protein Target Info (Source: Uniprot.org)

You can check the tissue specificity below for information on selecting positive and negative control.

Gene Name TNF
Protein Name Tumor necrosis factor
Molecular Weight 25644 MW
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). .
Sequence Similarities Belongs to the tumor necrosis factor family.
Subcellular Localization Cell membrane ; Single-pass type II membrane protein .
Uniprot ID P01375
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;
Research Areas |immunology|innate immunity|cytokines|tnf superfamily| signal transduction|growth factors/hormones|tnf| cancer|growth factors| cardiovascular|atherosclerosis|vascular inflammation|inflammatory mediators| metabolism|types of disease|metabolic disorders|
*if product is indicated to react with multiple species, protein info is based on the human gene.

Background for Tumor necrosis factor

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.

Human TNF Alpha PicoKine™ ELISA Kit Images

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Human TNF Alpha PicoKine™ ELISA Kit
Human TNF alpha PicoKine ELISA Kit standard curve

Intra/Inter Assay Precision

>
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n161616242424
Mean(pg/ml)93327608102319613
Standard deviation5.115.4317.6515.335
CV(%)5.54.75.17.54.85.7

Typical Data Obtained from Human TNF Alpha PicoKine™ ELISA Kit

(TMB reaction incubate at 37°C for 20-25min)

Concentration(pg/ml)015.631.262.51252505001000
O.D.0.0650.1180.1670.2720.5100.8391.5822.412

*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.

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Publications

Du, X., Dong, B., Li, C., Zhang, F., Ji, Y., Zhang, J., & Yin, C. (2017). Dynamic changes of α-melanocyte-stimulating hormone levels in the serum of patients with craniocerebral trauma. Experimental and Therapeutic Medicine, 14(3), 2511-2516. doi: 10.3892/etm.2017.4793
Ballerini, P., Diomede, F., Petragnani, N., Cicchitti, S., Merciaro, I., Cavalcanti, M.F.X.B., & Trubiani, O. (2017). Conditioned medium from relapsing-remitting multiple sclerosis patients reduces the expression and release of inflammatory cytokines induced by LPS-gingivalis in THP-1 and MO3.13 cell lines. Cytokine, 96, 261-272. Advance online publication. doi: 10.1016/j.cyto.2017.04.022
Choghakhori, R., Abbasnezhad, A., Hasanvand, A., & Amani, R. (2017). Inflammatory cytokines and oxidative stress biomarkers in irritable bowel syndrome: Association with digestive symptoms and quality of life. Cytokine. Advance online publication. doi: 10.1016/j.cyto.2017.05.005
Sun H, Zhu X, Cai W, Qiu L. Sun H1, Zhu X2, Cai W3, Qiu L4. Hypaphorine Attenuates Lipopolysaccharide-Induced Endothelial Inflammation via Regulation of TLR4 and PPAR-γ Dependent on PI3K/Akt/mTOR Signal Pathway.
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 urkish Journal of Biochemistry. 20160168, ISSN (Online) 1303-829X, ISSN (Print) 0250-4685, DOI: https://doi.org/10.1515/tjb-2016-0168, January 2017 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 THE EGYPTIAN JOURNAL OF IMMUNOLOGY Vol. 24 (1), 2017 Page: 09-20 IL-17 Induced The Recruitment and Functional Activity of Granulocytes Isolated from Patients Coinfected with Schistosoma mansoni and Hepatitis C Virus
Zijian Zhoua, b, Bin Lua, b, Chen Wanga, b, Zongqi Wanga, b, Tianfei Luob, c, Meihua Piaod, Fankai Menge, Guangfan Chif, Yinan Luoa, Pengfei Ge Cancer Letters Volume 390, 1 April 2017, Pages 77–90 RIP1 and RIP3 contribute to shikonin-induced DNA double-strand breaks in glioma cells via increase of intracellular reactive oxygen species
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 Accepted manuscript online: 18 January 2017 DOI: 10.1111/jpi.1238 Melatonin alleviates cadmium-induced liver injury by inhibiting the TXNIP-NLRP3 inflammasome
Jianzhong Yang, Mayra Azat, Peng Peng, Yiping Chang, Ranran Gao, Wenli Li, Jiming Li, Yage Chai, Dandan Li Int J Clin Exp Pathol 2017;10(1):781-788 www.ijcep.com /ISSN:1936-2625/IJCEP0033609 Serum levels of TNF-α, IL-1β, IL-9, and IL-15 in acute respiratory distress syndrome
Zhao, B., Pan, Y., Xu, H. et al. J Headache Pain (2017) 18: 1. doi:10.1186/s10194-016-0713-y Hyperbaric oxygen attenuates neuropathic pain and reverses inflammatory signaling likely via the Kindlin-1/Wnt-10a signaling pathway in the chronic pain injury model in rats
Jia Liu, Qianxue Chen, Zhihong Jian, Xiaoxing Xiong, Lingmin Shao, Tong Jin, Xiqun Zhu, and Lei Wang Department of Neurosurgery, Renmin Hospital of Wuhan University, Wuhan, Hubei 430060, China Daphnetin Protects against Cerebral Ischemia/Reperfusion Injury in Mice via Inhibition of TLR4/NF-κB Signaling Pathway
Wen-Juan Li, xiao-fang Tang, Xiao-Xue Shuai, Cheng-Jia Jiang, Xiang Liu, Le-Feng Wang, Yu-Fei Yao, Shaoping Nie, and Ming-Yong Xie J Agric Food Chem. 2016 Dec 8. [Epub ahead of print] Mannose receptor mediates the immune response to Ganoderma atrum polysaccharides in macrophages
JIANSHENG WANG, BAISHAN SU, HONGBIN ZHU, CHAO CHEN and GANG ZHAO Department of Orthopedics, The Second Hospital of Tangshan, Tangshan, Hebei 063000, P.R. China Received April 25, 2015; Accepted July 26, 2016 DOI: 10.3892/etm.2016.3850 Protective effect of geraniol inhibits inflammatory response, oxidative stress and apoptosis in traumatic injury of the spinal cord through modulation of NF-κB and p38 MAPK
Duan Y, Gu X, Zhu D, Sun W, Chen J, Feng J, Song K, Xu F, He X, He X. Int J Parasitol. 2014 Mar;44(3-4):217-24. Doi: 10.1016/J.Ijpara.2013.11.003. Epub 2014 Jan 31. Schistosoma Japonicum Soluble Egg Antigens Induce Apoptosis And Inhibit Activation...
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Zhang S, Liu X, Goldstein S, Li Y, Ge J, He B, Fei X, Wang Z, Ruiz G. Mol Med Rep. 2013 Jan;7(1):93-8. Doi: 10.3892/Mmr.2012.1159. Epub 2012 Oct 29. Role Of The Jak/Stat Signaling Pathway In The Pathogenesis Of Acute Myocardial Infarction In Rats ...
Wang J, Wang Z, Yao Y, Wu J, Tang X, Gu T, Li G. Tuberculosis (Edinb). 2015 Jul;95(4):505-14. Doi: 10.1016/J.Tube.2015.04.006. Epub 2015 May 9. The Fibroblast Growth Factor-2 Arrests Mycobacterium Avium Sp. Paratuberculosis Growth And Immunomodula...
Zhang Hq, Ding Tt, Zhao Js, Yang X, Zhang Hx, Zhang Jj, Cui Yl. World J Gastroenterol. 2009 Apr 21;15(15):1821-8. Therapeutic Effects Of Clostridium Butyricum On Experimental Colitis Induced By Oxazolone In Rats.
Chen J, Chen X, Qin J. Brain Inj. 2011;25(3):292-9. Doi: 10.3109/02699052.2010.546824. Epub 2011 Jan 31. Effects Of Polysaccharides Of The Euphoria Longan (Lour.) Steud On Focal Cerebral Ischemia/Reperfusion Injury And Its Underlying Mechanism.
Wang W, Tang L, Li Y, Wang Y. J Neurol Sci. 2015 Jan 15;348(1-2):121-5. Doi: 10.1016/J.Jns.2014.11.018. Epub 2014 Nov 18. Biochanin A Protects Against Focal Cerebral Ischemia/Reperfusion In Rats Via Inhibition Of P38-Mediated Inflammatory Responses.
Li J, Zhao L, He X, Zeng Yj, Dai Ss. Plos One. 2013;8(3):E59257. Doi: 10.1371/Journal.Pone.0059257. Epub 2013 Mar 15. Sinomenine Protects Against Lipopolysaccharide-Induced Acute Lung Injury In Mice Via Adenosine A(2A) Receptor Signaling.
Alikhani M, Khalighinejad N, Ghalaiani P, Khaleghi Ma, Askari E, Gorsky M. Oral Surg Oral Med Oral Pathol Oral Radiol. 2014 Jul;118(1):68-71. Doi: 10.1016/J.Oooo.2014.03.007. Epub 2014 Mar 15. Immunologic And Psychologic Parameters Associated With...
Zhang Z, Wu Y, Zhao Y, Xiao X, Liu J, Zhou X. Exp Ther Med. 2013 May;5(5):1523-1527. Epub 2013 Mar 22. Dynamic Changes In Hmgb1 Levels Correlate With Inflammatory Responses During Cardiopulmonary Bypass.
Wang D, Zhu X, Cui C, Dong M, Jiang H, Li Z, Liu Z, Zhu W, Wang Jg. J Chem Inf Model. 2013 Feb 25;53(2):343-53. Doi: 10.1021/Ci3004545. Epub 2013 Jan 30. Discovery Of Novel Acetohydroxyacid Synthase Inhibitors As Active Agents Against Mycobacteriu...
Shen B, Li J, Gao L, Zhang J, Yang B. Mol Cell Biochem. 2013 Jun;378(1-2):137-44. Doi: 10.1007/S11010-013-1604-Z. Epub 2013 Mar 3. Role Of Cc-Chemokine Receptor 5 On Myocardial Ischemia-Reperfusion Injury In Rats.
Yue P, Rong X, Zhuang X, Sha Hj, Li Jm, Xin L, Li Qw. Fish Physiol Biochem. 2014 Apr;40(2):625-34. Doi: 10.1007/S10695-013-9871-X. Epub 2013 Oct 25. Cloning And Expression Analysis Of A Novel High-Mobility Group Box 2 Homologue From Lampetra Japon...
Zhu G, Du Q, Wang X, Tang N, She F, Chen Y. Int J Mol Med. 2014 Jun;33(6):1431-40. Doi: 10.3892/Ijmm.2014.1711. Epub 2014 Mar 24. Tnf-?? Promotes Gallbladder Cancer Cell Growth And Invasion Through Autocrine Mechanisms.
Qin T, Yin Y, Yu Q, Yang Q. Plos One. 2015 Feb 6;10(2):E0117477. Doi: 10.1371/Journal.Pone.0117477. Ecollection 2015. Bursopentin (Bp5) Protects Dendritic Cells From Lipopolysaccharide-Induced Oxidative Stress For Immunosuppression.
Wang Yp, Wu Y, Li Ly, Zheng J, Liu Rg, Zhou Jp, Yuan Sy, Shang Y, Yao Sl. J Neuroinflammation. 2011 Aug 10;8:95. Doi: 10.1186/1742-2094-8-95. Aspirin-Triggered Lipoxin A4 Attenuates Lps-Induced Pro-Inflammatory Responses By Inhibiting Activation O...
Peng J, Wang P, Ge H, Qu X, Jin X. Plos One. 2015 May 1;10(5):E0125902. Doi: 10.1371/Journal.Pone.0125902. Ecollection 2015. Effects Of Cordycepin On The Microglia-Overactivation-Induced Impairments Of Growth And Development Of Hippocampal Culture...
Song Y, Chen X, Wang Ly, Gao W, Zhu Mj. Cns Neurosci Ther. 2013 Aug;19(8):603-10. Doi: 10.1111/Cns.12116. Epub 2013 May 3. Rho Kinase Inhibitor Fasudil Protects Against ??-Amyloid-Induced Hippocampal Neurodegeneration In Rats.
Zhang X, Jin C, Li Y, Guan S, Han F, Zhang S. Food Chem Toxicol. 2013 Aug;58:50-5. Doi: 10.1016/J.Fct.2013.04.006. Epub 2013 Apr 21. Catalpol Improves Cholinergic Function And Reduces Inflammatory Cytokines In The Senescent Mice Induced By D-Galac...
Zhang L, Ya B, Yang P, Sun F, Zhang L, Li Y, Li L. Neurochem Res. 2014 Apr;39(4):653-60. Doi: 10.1007/S11064-014-1242-1. Epub 2014 Jan 29. Impact Of Carotid Atherosclerosis Combined With Hypercholesterolemia On Cerebral Microvessels And Brain Pare...
Song Q, Jiang Z, Li N, Liu P, Liu L, Tang M, Cheng G. Biomaterials. 2014 Aug;35(25):6930-40. Doi: 10.1016/J.Biomaterials.2014.05.002. Epub 2014 May 27. Anti-Inflammatory Effects Of Three-Dimensional Graphene Foams Cultured With Microglial Cells.
Cheleschi S, Cantarini L, Pascarelli Na, Collodel G, Lucherini Om, Galeazzi M, Fioravanti A. Cytokine. 2015 Feb;71(2):165-72. Doi: 10.1016/J.Cyto.2014.10.023. Epub 2014 Nov 17. Possible Chondroprotective Effect Of Canakinumab: An In Vitro Study On...
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Zahedi M, Ghiasvand R, Feizi A, Asgari G, Darvish L. Int J Prev Med. 2013 Jul;4(7):777-85. Does Quercetin Improve Cardiovascular Risk Factors And Inflammatory Biomarkers In Women With Type 2 Diabetes: A Double-Blind Randomized Controlled Clinical ...
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Hara T, Mimura K, Abe T, Shioi G, Seiki M, Sakamoto T. J Biol Chem. 2011 Sep 16;286(37):32542-51. Doi: 10.1074/Jbc.M111.271726. Epub 2011 Jul 21. Deletion Of The Mint3/Apba3 Gene In Mice Abrogates Macrophage Functions And Increases Resistance To L...
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Chen T, Liu W, Chao X, Zhang L, Qu Y, Huo J, Fei Z. Brain Res Bull. 2011 Feb 1;84(2):163-8. Doi: 10.1016/J.Brainresbull.2010.11.015. Epub 2010 Dec 4. Salvianolic Acid B Attenuates Brain Damage And Inflammation After Traumatic Brain Injury In Mice.
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Xia J, Zhou Xl, Zhao Y, Zhu Yq, Jiang S, Ni Sz. Inflammation. 2013 Dec;36(6):1431-9. Doi: 10.1007/S10753-013-9683-2. Roles Of Lipoxin A4 In Preventing Paracetamol-Induced Acute Hepatic Injury In A Rabbit Model.
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FAQs

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