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Human IL-1 Beta PicoKine™ ELISA Kit
| SKU | EK0392 |
|---|---|
| Size | 96wells/kit, with removable strips. |
| Reactivity | Human |
| Sample Type | cell culture supernates, serum and plasma(heparin, EDTA, citrate). |
| Sensitivity | <0.15pg/ml |
| Assay Range | 3.9pg/ml-250pg/ml |
Overview
| Product Name | Human IL-1 Beta PicoKine™ ELISA Kit |
|---|---|
| SKU/Catalog Number | EK0392 |
| 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 IL-1 beta. 96wells/kit, with removable strips. |
| Cite This Product | Human IL-1 Beta PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0392) |
| 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 thata are not specified above or contact us to check for feasibility. |
| Sensitivity | <0.15pg/ml |
| Assay Range | 3.9pg/ml-250pg/ml |
| Immunogen | Expression system for standard: E.coli,A117-S269 |
| Reactivity | Human |
| Cross Reactivity | There is no detectable cross-reactivity with other relevant proteins. |
Assay Details
Kit Components
| Catalog number | Description | Quantity |
| EK0392-CAP | 96-well plate precoated with anti-Human IL1B antibody | 1 |
| EK0392-ST | lyophilized recombinant Human IL1B standard | 1ng/tube |
| EK0392-DA | biotinylated anti-Human IL1B antibody | 130ul(dilution 1:100) |
| AR1103 | Avidin-Biotin-Peroxidase Complex(ABC) | 130ul(dilution 1:100) |
| 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 IL-1 Beta PicoKine™ ELISA Kit
(TMB reaction incubate at 37°C for 15-20min)
| Concentration(pg/ml) | 0 | 3.9 | 7.8 | 15.6 | 31.2 | 62.5 | 125 | 250 |
| O.D. | 0.026 | 0.104 | 0.176 | 0.319 | 0.577 | 1.033 | 1.765 | 2.466 |
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) | 11 | 60 | 123 | 10 | 58 | 112 |
| Standard deviation | 0.45 | 2.88 | 8.97 | 0.57 | 3.24 | 9.96 |
| CV(%) | 4.1% | 4.8% | 7.3% | 5.7% | 5.6% | 8.9% |
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 | 11 | 10 | 11 | 11 | 10 | 0.43 | 4.3% |
| Sample 2 | 60 | 54 | 52 | 53 | 54 | 3.11 | 5.7% |
| Sample 3 | 123 | 119 | 123 | 112 | 119 | 4.49 | 3.7% |
*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 | P01584 |
|---|---|
| Gene Name | IL1B |
| Protein Name | Interleukin-1 beta |
| Alternative Names | Interleukin-1 beta;IL-1 beta;Catabolin;IL1B;IL1F2; |
| 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 | 30748 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. . |
|---|---|
| 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 alone. 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 genes. Furthermore, Microenvironmental IL-1beta and, to a lesser extent, IL-1alpha are required for in vivo angiogenesis and invasiveness of different tumor cells. Additional, the cooperation of IL-1beta and PDGFB induces contractile-to-synthetic phenotype modulation of human aortic smooth muscle cells in culture. 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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Customer Reviews
- Review by Dhruva Bhattacharya
-
Rating: (Posted on 12/21/2017)
Source: Biocompare.com
"User Friendly And Economical ELISA Kit"
--Dhruva Bhattacharya, University of Arizona, Opthalmology and Vision Science, Postdoctoral Research Associate
Application ELISA Sample "Culture media supernatant" of transformed cultured cells of human conjunctiva epithelial cells Detection -- "I am very much satisfied with the product Human IL-1 Beta PicoKine™ ELISA Kit (EK0392) purchased from Boster Bio. I used the Boster Bio ElISA kit for the first time and I am very happy with this product. After trying the IL-1beta kit, I have purchased two other cytokine detection ELISA kits from Boster Bio. The methodology of Human IL-1 Beta PicoKine™ ELISA Kit is simplified and user friendly. It required minimum optimization for my experiments. The written instructions provided with the kit is very useful for running the ELISA. The results obtained so far have been consistent and reproducible. For my future ELISA experiments, I'll be buying the ELISA kit from Boster Bio."
- Review by Milos Lazic
-
Rating: (Posted on 8/26/2016)
Source: Biocompare.com
"Good quality budget option"
--Milos Lazic, JECURE THERAPEUTICS, SCIENTIST II
Applications ELISA Sample protein concentrations in cell culture media Detection -- "We are a pharmaceutical startup optimizing cell based drug assays where one of our endpoints was observing the differences in mature IL-1b production in the media. I have compared several kits from the most expensive versions of R&D, over eBioscience/Affymetrix to a more budget Boster's kit. The aim was to decide what is the best price for the value and quality we are getting for reliable and replicable ELISA analyses. This assay was used to test the effect of increasing concentrations of a drug on IL-1b levels (dose response). The kit reported similar data obtained from the other two kits (R&D and eBiosciences), with a slightly larger standard deviations. Results replicated the findings of the previous kits and the assay had good sensitivity/ instructions on how to perform assay and analyze data. I would use this assay in an academic lab for a project where no more than 2-3 96-well plates will be used. It should be done carefully by a scientist that has previously performed ELISA assays. There are many steps to it. It is a good quality budget option to the other leading kits on the market. I would not recommend it for a larger drug screen assay where more than 5 ELISAs kits ought to be used."
Publications
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
Wan L, Yuan X, Liu M, Xue B. Exp Ther Med. 2018 Mar;15(3):2429-2435. doi: 10.3892/etm.2017.5667. Epub 2017 Dec 21. miRNA-223-3p regulates NLRP3 to promote apoptosis and inhibit proliferation of hep3B cells
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
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
Qin Q, Xu G, Qi W, Guo M, Wang Z, Xu W, Qiao Z, Gu Y, Niu J. Exp Ther Med. 2017 Aug;14(2):1017-1022. doi: 10.3892/etm.2017.4609. Epub 2017 Jun 15. Evaluation of the C-domain of heparanase during AGE-induced macrophage inflammatory response
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
Wang Y, Qin Y, Wang T, Chen Y, Lang X, Zheng J, Gao S, Chen S, Zhong X, Mu Y, Wu X, Zhang F, Zhao W, Zhong Z. Sci Rep. 2018 Feb 13;8(1):2887. doi: 10.1038/s41598-018-20958-1. Pyroptosis induced by enterovirus 71 and coxsackievirus B3 infection affects viral replication and host response
Tâlvan, E.T., Mohor, C., Chisnoiu, D., Cristea, V., & Câmpian, R.S. (2017). Expression of Interleukin (IL)-1β, IL-8, IL-10 and IL-13 in Chronic Adult Periodontitis Progression. Archives of Medicine, 9(3:4), 1-8. doi: 10.21767/1989-5216.1000219
El-Horany, H.E., Abd-Ellatif, R.N., Watany, M., Hafez, Y.M., Okda, H.I. (2017). NLRP3 expression and urinary HSP72 in relation to biomarkers of inflammation and oxidative stress in diabetic nephropathy patients. IUBMB Life. Advance online publication. doi: 10.1002/iub.1645
Jian, D., Dai, B., Hu, X., Yao, Q., Zhang, L., Zhu, J. (2017). ox-LDL increases microRNA-29a transcription through up-regulating YY1 and STAT1 in macrophages. Cell Biology International. Advance online publication. doi: 10.1002/cbin.10803
Tao, J., Zhang, D., Man, Y., Wang, W., & Bi, Y. (2017). Sodium ferulate inhibits high-fat diet-induced inflammatory factors expression in human umbilical vein endothelial cells. Cytology and Genetics, 51(3), 214-220. doi: 10.3103/S0095452717030124
Huang Y, Liu L, Ma D, Liao Y, Lu Y, Huang H, Qin W, Liu X, Fang F. Human cytomegalovirus triggers the assembly of AIM2 inflammasome in THP-1-derivedmacrophages. J Med Virol. 2017 May 8. doi: 10.1002/jmv.24846. [Epub ahead of print]
Jieliang Shen, Shengxi Xu, Hao Zhou, Huzhe Liu, Wei Jiang, Jie Hao & Zhenming Hu
Scientific Reports 7, Article number: 41067 (2017)
doi:10.1038/srep41067
IL-1β induces apoptosis and autophagy via mitochondria pathway in human degenerative nucleus pulposus cells
Wang, HT., Huang, FL., Hu, ZL. et al. Neurotox Res (2017). doi:10.1007/s12640-016-9696-3
Early-Life Social Isolation-Induced Depressive-Like Behavior in Rats Results in Microglial Activation and Neuronal Histone Methylation that Are Mitigated by Minocycline
Jianbo Jia, Feifei Li, Shumei Zhai, Hongyu Zhou, Sijin Liu, Guibin Jiang, and Bing Yan
Environ. Sci. Technol., Just Accepted Manuscript
DOI: 10.1021/acs.est.6b05200
Publication Date (Web): January 9, 2017
Copyright © 2017 American Chemical Society
Susceptibility of Overweight Mice to Liver Injury as a Result of the ZnO Nanoparticle-Enhanced Liver Deposition of Pb2+
Structural characterization and immunostimulatory activity of a glucan from natural Cordyceps sinensis
Junqiao Wanga, Shaoping Niea, , , Steve W. Cuia, b, Zhijun Wanga, Aled O. Phillipsc, Glyn O. Phillipsd, Yajing Lie, Mingyong Xiea,
Structural characterization and immunostimulatory activity of a glucan from natural Cordyceps sinensis
G Xu, Q Qin, M Yang, Z Qiao, Y Gu, J Niu - Diabetes Research and Clinical Practice, 2016
Heparanase-driven Inflammation from the AGEs-stimulated Macrophages Changes the Functions of Glomerular Endothelial Cells
Janahmadi, Z., Nekooeian, A.A., Moaref, A.R. et al. Naunyn-Schmiedeberg's Arch Pharmacol (2016). doi:10.1007/s00210-016-1323-6
Oleuropein attenuates the progression of heart failure in rats by antioxidant and antiinflammatory effects
Customer Q&As
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: 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 human il 1b elisa kit, human il 1 beta elisa kit, human il 1 elisa kit, human il1 beta elisa kit, human il-1 beta elisa kit, human il1b elisa kit