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| SKU | EK0394 |
|---|---|
| Size | 96wells/kit, with removable strips. |
| Reactivity | Mouse |
| Sample Type | cell culture supernates, cell lysates, serum and plasma (heparin, EDTA). |
| Sensitivity | <1pg/ml |
| Assay Range | 12.5pg/ml-800pg/ml |
Overview
| Product Name | Mouse IL-1 Beta PicoKine™ ELISA Kit |
|---|---|
| SKU/Catalog Number | EK0394 |
| 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 Mouse IL-1 beta. 96wells/kit, with removable strips. |
| Cite This Product | Mouse IL-1 Beta PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0394) |
| Sample Type | cell culture supernates, cell lysates, serum and plasma (heparin, EDTA)..
Anticoagulant(s): heparin or EDTA
*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 | 12.5pg/ml-800pg/ml |
| Immunogen | Expression system for standard: E.coli,V118-S269 |
| Reactivity | Mouse |
| Cross Reactivity | There is no detectable cross-reactivity with other relevant proteins. |
| Antibody Clonalities | Capture Antibody | Detection Antibody: monoclonal antibody from rat|polyclonal antibody from goat |
Assay Details
Kit Components
| Catalog number | Description | Quantity |
| EK0394-CAP | 96-well plate precoated with anti-Mouse IL1B antibody | 1 |
| EK0394-ST | lyophilized recombinant Mouse IL1B standard | 10ng/tube |
| EK0394-DA | biotinylated anti-Mouse IL1B 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 Mouse IL-1 Beta PicoKine™ ELISA Kit
(TMB reaction incubate at 37°C for 15-25min)
| Concentration(pg/ml) | 0 | 12.5 | 25 | 50 | 100 | 200 | 400 | 800 |
| O.D. | 0.058 | 0.100 | 0.143 | 0.215 | 0.420 | 0.725 | 1.362 | 2.135 |
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) | 33 | 211 | 483 | 36 | 217 | 481 |
| Standard deviation | 1.74 | 10.33 | 34.29 | 2.19 | 14.75 | 35.59 |
| CV(%) | 5.3% | 4.9% | 7.1% | 6.1% | 6.8% | 7.4% |
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 | 33 | 34 | 31 | 32 | 32 | 1.11 | 3.4% |
| Sample 2 | 211 | 206 | 203 | 208 | 207 | 2.91 | 1.4% |
| Sample 3 | 483 | 452 | 433 | 430 | 449 | 21.1 | 4.6% |
*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 | P10749 |
|---|---|
| Gene Name | IL1B |
| Protein Name | Interleukin-1 beta |
| Alternative Names | Interleukin-1 beta;IL-1 beta;Il1b; |
| Subcellular Localization | Secreted. The lack of a specific hydrophobic segment in the precursor sequence suggests that IL-1 is released by damaged cells or is secreted by a mechanism differing from that used for other secretory proteins. |
| Molecular Weight | 30931 MW |
*if product is indicated to react with multiple species, protein info is based on the human gene.
Ontology
| Protein Function | Produced by activated macrophages, IL-1 stimulates thymocyte proliferation by inducing IL-2 release, B-cell maturation and proliferation, and fibroblast growth factor activity. IL-1 proteins are involved in the inflammatory response, being identified as endogenous pyrogens, and are reported to stimulate the release of prostaglandin and collagenase from synovial cells. |
|---|---|
| Research Areas | Atherosclerosis, Cardiovascular, Cytokines, Immunology, Innate Immunity, Interleukins, Metabolism, Obesity, Vascular Inflammation *You can search these to find other products in these research areas. |
| Background | Interleukin-1beta(IL-1beta) is a potent stimulator of bone resorption whose gene is mapped to 2q14, and has been implicated in the pathogenesis of high bone turnover and osteoporosis. IL-1beta, a prominent microglia-derived cytokine, caused oligodendrocyte death in coculture with astrocytes and microglia, but not in pure culture of oligodendrocytes alone1. It also can cause nuclear export of a specific NCOR corepressor complex, resulting in derepression of a specific subset of nuclear factor-kappa-B(NFKB)-regulated genes2. Furthermore, Microenvironmental IL-1beta and, to a lesser extent, IL-1alpha are required for in vivo angiogenesis and invasiveness of different tumor cells3. Additional, the cooperation of IL-1beta and PDGFB induces contractile-to-synthetic phenotype modulation of human aortic smooth muscle cells in culture4. Moreover, the association with disease may be explained by the biologic properties of IL-1beta, which is an important proinflammatory cytokine and a powerful inhibitor of gastric acid secretion. |
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Publications
Ascorbic acid ameliorates oxidative stress and inflammation in dextran sulfate sodium-induced ulcerative colitis in mice.
Death-associated protein kinase 1 mediates interleukin-1? production through regulating inlfammasome activation in Bv2 microglial cells and mice
Grape seed proanthocyanidin extract supplementation affects exhaustive exercise-induced fatigue in mice
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Pre-B cell leukemia transcription factor 3 induces inflammatory responses in human umbilical vein endothelial cells and murine sepsis via acting a competing endogenous RNA for high mobility group box 1 protein
Luteolin alleviates NLRP3 inflammasome activation and directs macrophage polarization in lipopolysaccharide-stimulated RAW264.7 cells
Nicotine promotes atherosclerosis via ROS-NLRP3-mediated endothelial cell pyroptosis
Toxoplasma gondii excretory/secretory antigens (TgESAs) suppress pro-inflammatory cytokine secretion by inhibiting TLR-induced NF-?B activation in LPS-stimulated murine macrophages
VSIG4 inhibits proinflammatory macrophage activation by reprogramming mitochondrial pyruvate metabolism
TLR2/TLR4 activation induces Tregs and suppresses intestinal inflammation caused by Fusobacterium nucleatum in vivo
Tauroursodeoxycholic acid attenuates endoplasmic reticulum stress and protects the liver from chronic intermittent hypoxia induced injury
Oroxylin A inhibits colitis by inactivating NLRP3 inflammasome
Effects of Recombinant Toxoplasma gondii Citrate Synthase I on the Cellular Functions of Murine Macrophages In vitro
Evaluation of the chemical composition, antioxidant and anti-inflammatory activities of distillate and residue fractions of sweet basil essential oil
Liver X Receptor Agonist TO901317 Attenuates Paraquat-Induced Acute Lung Injury through Inhibition of NF-?B and JNK/p38 MAPK Signal Pathways
Foxo3a-dependent Bim transcription protects mice from a high fat diet via inhibition of activation of the NLRP3 inflammasome by facilitating autophagy flux in Kupffer cells
High-Throughput Sequencing and Co-Expression Network Analysis of lncRNAs and mRNAs in Early Brain Injury Following Experimental Subarachnoid Haemorrhage
Oral Administration of Resveratrol Alleviates Osteoarthritis Pathology in C57BL/6J Mice Model Induced by a High-Fat Diet
Ginsenoside Rg1 improves fertility and reduces ovarian pathological damages in premature ovarian failure model of mice
Heparanase Mediates Intestinal Inflammation and Injury in a Mouse Model of Sepsis
Daphnetin Protects against Cerebral Ischemia/Reperfusion Injury in Mice via Inhibition of TLR4/NF-?B Signaling 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
Sulfated Cyclocarya paliurus polysaccharides markedly attenuates inflammation and oxidative damage in lipopolysaccharide-treated macrophage cells and mice
Chinese herb cinobufagin-reduced cancer pain is associated with increased peripheral opioids by invaded CD3/4/8 lymphocytes
NEMO-Binding Domain Peptide Attenuates Lipopolysaccharide-Induced Acute Lung Injury by Inhibiting the NF-?B Signaling Pathway
LL202 protects against dextran sulfate sodium-induced experimental colitis in mice by inhibiting MAPK/AP-1 signaling
Potential implications of Apolipoprotein E in early brain injury after experimental subarachnoid hemorrhage: Involvement in the modulation of blood-brain barrier integrity
Nuclear Factor E2-Related Factor-2 Negatively Regulates NLRP3 Inflammasome Activity by Inhibiting Reactive Oxygen Species-Induced NLRP3 Priming
Synergistic Effect of Compounds from a Chinese Herb: Compatibility and Dose Optimization of Compounds from N-Butanol Extract of Ipomoea stolonifera
PER1 prevents excessive innate immune response during endotoxin-induced liver injury through regulation of macrophage recruitment in mice
Chemopreventive activity of GEN-27, a genistein derivative, in colitis-associated cancer is mediated by p65-CDX2-?-catenin axis
Celastrol Ameliorates Ulcerative Colitis-Related Colorectal Cancer in Mice via Suppressing Inflammatory Responses and Epithelial-Mesenchymal Transition
Unfractionated heparin attenuates intestinal injury in mouse model of sepsis by inhibiting heparanase
Homocysteine Triggers Inflammatory Responses in Macrophages through Inhibiting CSE-H2S Signaling via DNA Hypermethylation of CSE Promoter
Effects of Cordycepin on the Microglia-Overactivation-Induced Impairments of Growth and Development of Hippocampal Cultured Neurons
Changes of cytokine levels in a mouse model of post-infectious irritable bowel syndrome
CYP epoxygenase 2J2 prevents cardiac fibrosis by suppression of transmission of pro-inflammation from cardiomyocytes to macrophages
Minocycline treatment ameliorates interferon-alpha- induced neurogenic defects and depression-like behaviors in mice
Interleukin-22 ameliorates liver fibrogenesis by attenuating hepatic stellate cell activation and downregulating the levels of inflammatory cytokines
Epoxyeicosatrienoic Acids Regulate Macrophage Polarization and Prevent LPS-Induced Cardiac Dysfunction
Toll-like receptor 4 implicated in acute lung injury induced by paraquat poisoning in mice
Antenatal exposure of maternal secondhand smoke (SHS) increases fetal lung expression of RAGE and induces RAGE-mediated pulmonary inflammation
Agmatine Protects against Zymosan-Induced Acute Lung Injury in Mice by Inhibiting NF-?B-Mediated Inflammatory Response
Anti-Inflammatory Activity of N-Butanol Extract from?Ipomoea stolonifera In Vivo?and?In Vitro
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Astragalus membranaceus Inhibits Inflammation via Phospho-P38 Mitogen-Activated Protein Kinase (MAPK) and Nuclear Factor (NF)-?B Pathways in Advanced Glycation End Product-Stimulated Macrophages
The NALP3 inflammasome is involved in neurotoxic prion peptide-induced microglial activation
Aspirin-triggered lipoxin A4?attenuates LPS-induced pro-inflammatory responses by inhibiting activation of NF-?B and MAPKs in BV-2 microglial cells
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Li J, Diao B, Guo S, Huang X, Yang C, Feng Z, Yan W, Ning Q, Zheng L, Chen Y, Wu Y. Nat Commun. 2017 Nov 6;8(1):1322. doi: 10.1038/s41467-017-01327-4. VSIG4 inhibits proinflammatory macrophage activation by reprogramming mitochondrial pyruvate metabolism
Jia YP, Wang K, Zhang ZJ, Tong YN, Han D, Hu CY, Li Q, Xiang Y, Mao XH, Tang B. PLoS One. 2017 Oct 9;12(10):e0186179. doi: 10.1371/journal.pone.0186179. eCollection 2017. TLR2/TLR4 activation induces Tregs and suppresses intestinal inflammation caused by Fusobacterium nucleatum in vivo
Liu X, Zhang X, Ding Y, Zhou W, Tao L, Lu P, Wang Y, Hu R. Antioxid Redox Signal. 2017 Jan 1;26(1):28-43. doi: 10.1089/ars.2015.6615. Epub 2016 Aug 9. Nuclear Factor E2-Related Factor-2 Negatively Regulates NLRP3 Inflammasome Activity by Inhibiting Reactive Oxygen Species-Induced NLRP3 Priming
Chen S, He Y, Hu Z, Lu S, Yin X, Ma X, Lv C, Jin G. J Histochem Cytochem. 2017 Apr;65(4):241-249. doi: 10.1369/0022155417692536. Epub 2017 Feb 7. Heparanase Mediates Intestinal Inflammation and Injury in a Mouse Model of Sepsis
Zhou W, Liu X, Zhang X, Tang J, Li Z, Wang Q, Hu R. Oncotarget. 2017 Jul 22;8(35):58903-58917. doi: 10.18632/oncotarget.19440. eCollection 2017 Aug 29. Oroxylin A inhibits colitis by inactivating NLRP3 inflammasome
Lu Y, Zhang XS, Zhang ZH, Zhou XM, Gao YY, Liu GJ, Wang H, Wu LY, Li W, Hang CH. J Neuroinflammation. 2018 Mar 19;15(1):87. doi: 10.1186/s12974-018-1118-4. Peroxiredoxin 2 activates microglia by interacting with Toll-like receptor 4 after subarachnoid hemorrhage
Zhang Y, Feng J, Cui J, Yang G, Zhu X. Mol Med Rep. 2018 Apr;17(4):5805-5813. doi: 10.3892/mmr.2018.8609. Epub 2018 Feb 15. Pre-B cell leukemia transcription factor 3 induces inflammatory responses in human umbilical vein endothelial cells and murine sepsis via acting a competing endogenous RNA for high mobility group box 1 protein
Wu X, Zhang H, Qi W, Zhang Y, Li J, Li Z, Lin Y, Bai X, Liu X, Chen X, Yang H, Xu C, Zhang Y, Yang B. Cell Death Dis. 2018 Feb 7;9(2):171. doi: 10.1038/s41419-017-0257-3. Nicotine promotes atherosclerosis via ROS-NLRP3-mediated endothelial cell pyroptosis
Zhang BC, Li Z, Xu W, Xiang CH, Ma YF Am J Transl Res. 2018 Jan 15;10(1):265-273. eCollection 2018. Luteolin alleviates NLRP3 inflammasome activation and directs macrophage polarization in lipopolysaccharide-stimulated RAW264.7 cells
Du, J., Chen, X., Wang, C., & Sun, H. (2017). Pathway analysis of global gene expression change in dendritic cells induced by the polysaccharide from the roots of Actinidia eriantha. Journal of Ethnopharmacology. Advance online publication. doi: 10.1016/j.jep.2017.12.009
Wang, Q., Dong, X., Wang, Y., Liu, M., Sun, A., Li, N.,..., & Li, X. (2017). Adolescent escitalopram prevents the effects of maternal separation on depression- and anxiety-like behaviours and regulates the levels of inflammatory cytokines in adult male mice. International Journal of Developmental Neuroscience, 62, 37-45. doi: 10.1016/j.ijdevneu.2017.07.007
Feng, T., Wei, S., Wang, Y., Fu, X., Shi, L., Qu, L., & Fan, X. (2017). Rhein ameliorates adenomyosis by inhibiting NF-κB and β-Catenin signaling pathway. Biomedicine & Pharmacotherapy, 94, 231-237. doi: 10.1016/j.biopha.2017.07.089
Yan, J., Han, Z., Qu, Y., Yao, C., Shen, D., Tai, G.,..., & Zhou, Y. (2017). Structure elucidation and immunomodulatory activity of a β-glucan derived from the fruiting bodies of Amillariella mellea. Food Chemistry, 240, 534-543. doi: 10.1016/j.foodchem.2017.07.154
Zhang, M., Wu, X., Xu, Y., He, M., Yang, J., Li, J.,..., & Jia, J. (2017). The cystathionine β-synthase/hydrogen sulfide pathway contributes to microglia-mediated neuroinflammation following cerebral ischemia. Brain, Behavior, and Immunity. Advance online publication. doi: 10.1016/j.bbi.2017.07.156
Yu, Y., Shen, M., Wang, Z., Wang, Y., Xie, M., & Xie, J. (2017). Sulfated polysaccharide from Cyclocarya paliurusenhances the immunomodulatory activity of macrophages. Carbohydrate Polymers, 174, 669-676. doi: 10.1016/j.carbpol.2017.07.009
Dong, X.W., Jia, Y.L., Ge, L.T., Jiang, B., Jiang, J.X., Shen, J.,…, & Xie, Q.M. (2017). Soluble epoxide hydrolase inhibitor AUDA decreases bleomycin-induced pulmonary toxicity in mice by inhibiting the p38/Smad3 pathways. Toxicology, 389, 31-41. Advance online publication. doi: 10.1016/j.tox.2017.07.002
Zhang W, Li X, Liu Y, Chen H, Gong J. Biomed Pharmacother. 2017 Apr 19;90:821-834. doi: 10.1016/j.biopha.2017.04.025. Activation of imidazoline I1 receptor by moxonidine regulates the progression of liver fibrosis in the Nrf2-dependent pathway.
Wang Z, Xie J, Yang Y, Zhang F, Wang S, Wu T, Shen M, Xie M. Sci Rep. 2017 Jan 17;7:40402. doi: 10.1038/srep40402. Sulfated Cyclocarya paliurus polysaccharides markedly attenuates inflammation and oxidative damage in lipopolysaccharide-treated macrophage cells and mice
Ye Hh, Hua R, Yu L, Wu Kj, Fei Sj, Qin X, Song Y, Cao Jl, Zhang Ym. Dig Dis Sci. 2013 Oct;58(10):2826-39. Doi: 10.1007/S10620-013-2727-5. Epub 2013 Jun 7. Abnormal Expression Of Toll-Like Receptor 4 Is Associated With Susceptibility To Ethanol-Ind...
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 the TMB color developing agent contain H2O2?
A: Yes, the TMB color developing agent contain H2O2.
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: My plasma samples have been somewhat
hemolyzed. Does plasma hemolysis have any effect on ELISA results?
Keyword: application, protocol, troubleshooting
A: There will be some unspecific staining in hemolysis samples and make the result inaccurate. We suggest you collect plasma samples again, hemolysis samples are not recommended to use.
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: 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 mouse il 1b elisa kit, mouse il 1 beta elisa kit, mouse il 1 elisa kit, mouse il1 beta elisa kit, mouse il-1 beta elisa kit, mouse il1b elisa kit