| SKU: | EK0416 |
|---|---|
| Pack Size: | 96wells/kit, with removable strips. |
| Validated Species: | Human |
| Application: | ELISA |
| Sensitivity: | <0.5pg/ml |
| Assay Range: | 7.8pg/ml-500pg/ml(cellculturesupernates)3.4pg/ml-250pg/ml(humanserum,plasma) |
| Sample Type: | cell culture supernates, serum and plasma( heparin, EDTA, citrate). |
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Overview
| Product Name | Human IL-10 PicoKine™ ELISA Kit |
|---|---|
| SKU/Catalog Number | EK0416 |
| Description | Sandwich High Sensitivity ELISA kit for Quantitative Detection of Human IL-10. 96wells/kit, with removable strips. |
| Cite This Product | Human IL-10 PicoKine™ ELISA Kit (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0416) |
| 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; Immunogen sequence: S19-N178 |
| Cross Reactivity | There is no detectable cross-reactivity with other relevant proteins. |
| Pack Size | 96wells/kit, with removable strips. |
Properties
| Sensitivity | <0.5pg/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 | 7.8pg/ml-500pg/ml(cellculturesupernates)3.4pg/ml-250pg/ml(humanserum,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 | monoclonal antibody from mouse |
| Normal Levels | Some research articles suggesting the natural levels of IL10 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 number | Description | Quantity |
| EK0416-CAP | 96-well plate precoated with anti-Human IL10 antibody | 1 |
| EK0416-ST | lyophilized recombinant Human IL10 standard | 10ng/tubex2 |
| EK0416-DA | biotinylated anti-Human IL10 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 |
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 ComponentsMaterial 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.
Protein Target Info (Source: Uniprot.org)
You can check the tissue specificity below for information on selecting positive and negative control.
| Gene Name | IL10 |
|---|---|
| Protein Name | Interleukin-10 |
| Molecular Weight | 20517 MW |
| Protein Function | Inhibits the synthesis of a number of cytokines, including IFN-gamma, IL-2, IL-3, TNF and GM-CSF produced by activated macrophages and by helper T-cells. |
| Tissue Specificity | Produced by a variety of cell lines, including T-cells, macrophages, mast cells and other cell types. |
| Sequence Similarities | Belongs to the IL-10 family. |
| Subcellular Localization | Secreted. |
| Uniprot ID | P22301 |
| Alternative Names | Interleukin-10;IL-10;Cytokine synthesis inhibitory factor;CSIF;IL10; |
| Research Areas | |immunology|innate immunity|cytokines|interleukins| cancer|tumor immunology| immunology|adaptive immunity|regulatory t cells| |
Background for Interleukin-10
Interleukin-10, also called cytokine synthesis inhibitory factor, is implicated in tumorigenesis, and it has been shown that polymorphisms in its gene promoter correlate with differential amounts of production. IL-10 is an important cytokine with anti-inflammatory, anti-immune, and antifibrotic functions. It is also an important regulatory cytokine whose involvement extends into diverse areas of the human immune system. IL-10 is a recently described natural endogenous immunosuppressive cytokine that has been identified in human, murine, and other organisms. IL-10 significantly affects chemokine biology, because human IL-10 inhibits chemokine production and is a specific chemotactic factor for CD8+ T cells. It suppresses the ability of CD4+ T cells, but not CD8+ T cells, to migrate in response to IL-8. Interleukin-10 gene polymorphisms and interleukin-10 production capability may contribute to the development of skin squamous cell carcinomas after renal transplantation.1 The interleukin-10 locus contributes to the heritability of psoriasis susceptibility. With regard to sudden infant death, IL-10 is of special interest. This is an immunoregulatory cytokine that plays an important role in the development of infectious disease The mIL-10 gene is mapped to mouse chromosome 1 and the hIL-10 gene is also mapped to human chromosome 1. The standard product used in this kit is recombinant human IL-10, consisting of 160 amino acids with the molecular mass of 18.6KDa.
Human IL-10 PicoKine™ ELISA Kit Images
Click the images to enlarge.
Human IL-10 PicoKine ELISA Kit standard curve
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) | 25 | 112 | 280 | 34 | 184 | 312 |
| Standard deviation | 1.35 | 5.38 | 8.68 | 2.55 | 14.54 | 20.9 |
| CV(%) | 5.4 | 4.8 | 3.1 | 7.5 | 7.9 | 6.7 |
Typical Data Obtained from Human IL-10 PicoKine™ ELISA Kit
(TMB reaction incubate at 37°C for 15-20min)
| Concentration(pg/ml) | 0 | 7.8 | 15.6 | 31.3 | 62.5 | 125 | 250 | 500 | ||
| O.D. | 0.070 | 0.151 | 0.187 | 0.295 | 0.538 | 0.858 | 1.622 | 2.281 | ||
*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.
Publications
Dang, W., Xie, W. & Zhou, J. (2017). Expression of PYCARD Gene Transcript Variant mRNA in Peripheral Blood Mononuclear Cells of Primary Gout Patients with Different Chinese Medicine Syndromes. Chinese Journal of Integrative Medicine, 1-8. Advance online publication. doi: 10.1007/s11655-017-2826-9
He, S., Le, N.A., Frediani, J.K., Winterberg, P.D., Jin, R., Liverman, R.,..., & Vos, M.B. (2017). Cardiometabolic risks vary by weight status in pediatric kidney and liver transplant recipients: A cross-sectional, single-center study in the USA. Pediatric Transplantation, 21(6). doi: 10.1111/petr.12984
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
Zhang Y, Leng Yf, Xue X, Zhang Y, Wang T, Kang Yq. World J Gastroenterol. 2011 Jan 14;17(2):254-9. Doi: 10.3748/Wjg.V17.I2.254. Effects Of Penehyclidine Hydrochloride In Small Intestinal Damage Caused By Limb Ischemia-Reperfusion.
Luo H,, Wang X,, Wang J,, Chen C,, Wang N,, Xu Z,, Chen S,, Zeng C,. Cardiovasc Diabetol. 2015 Jun 10;14:76. Doi: 10.1186/S12933-015-0239-7. Chronic Nf-??b Blockade Improves Renal Angiotensin Ii Type 1 Receptor Functions And Reduces Blood Pressure ...
Wang T, Zhou Yt, Chen Xn, Zhu Ax, Wu Bh. World J Gastroenterol. 2014 Jul 28;20(28):9519-27. Doi: 10.3748/Wjg.V20.I28.9519. Remote Ischemic Postconditioning Protects Against Gastric Mucosal Lesions In Rats.
Zhang X, Liu F, Liu H, Cheng H, Wang W, Wen Q, Wang Y. Inflamm Res. 2011 Jun;60(6):569-75. Doi: 10.1007/S00011-010-0305-2. Epub 2011 Jan 19. Urinary Trypsin Inhibitor Attenuates Lipopolysaccharide-Induced Acute Lung Injury By Blocking The Activati...
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.
Li Xj, Liu Dp, Chen Hl, Pan Xh, Kong Qy, Pang Qf. Int Immunopharmacol. 2012 Feb;12(2):460-4. Doi: 10.1016/J.Intimp.2012.01.001. Epub 2012 Jan 15. Lactoferrin Protects Against Lipopolysaccharide-Induced Acute Lung Injury In Mice.
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.
Tang Y, Liao Y, Kawaguchi-Sakita N, Raut V, Fakhrejahani E, Qian N, Toi M. J Hepatobiliary Pancreat Sci. 2011 Jul;18(4):551-8. Doi: 10.1007/S00534-010-0368-Z. Sinisan, A Traditional Chinese Medicine, Attenuates Experimental Chronic Pancreatitis In...
Chen W, Qi J, Feng F, Wang Md, Bao G, Wang T, Xiang M, Xie Wf. Neurochem Int. 2014 Mar;68:28-37. Doi: 10.1016/J.Neuint.2014.01.015. Epub 2014 Feb 12. Neuroprotective Effect Of Allicin Against Traumatic Brain Injury Via Akt/Endothelial Nitric Oxide...
Ye Xh, Wu Y, Guo Pp, Wang J, Yuan Sy, Shang Y, Yao Sl. Brain Res. 2010 Apr 6;1323:174-83. Doi: 10.1016/J.Brainres.2010.01.079. Epub 2010 Feb 4. Lipoxin A4 Analogue Protects Brain And Reduces Inflammation In A Rat Model Of Focal Cerebral Ischemia R...
Jin Y, Wan Y, Chen G, Chen L, Zhang Mq, Deng L, Zhang Jc, Xiong Xz, Xin Jb. Plos One. 2014 Oct 16;9(10):E111044. Doi: 10.1371/Journal.Pone.0111044. Ecollection 2014. Treg/Il-17 Ratio And Treg Differentiation In Patients With Copd.
Sereshki N, Gharagozloo M, Ostadi V, Ghahiri A, Roghaei Ma, Mehrabian F, Andalib Aa, Hassanzadeh A, Hosseini H, Rezaei A. Int J Fertil Steril. 2014 Apr;8(1):59-66. Epub 2014 Mar 9. Variations In T-Helper 17 And Regulatory T Cells During The Menstr...
Hualin C, Wenli X, Dapeng L, Xijing L, Xiuhua P, Qingfeng P. Inflammation. 2012 Jun;35(3):1087-93. Doi: 10.1007/S10753-011-9415-4. The Anti-Inflammatory Mechanism Of Heme Oxygenase-1 Induced By Hemin In Primary Rat Alveolar Macrophages.
Zhou X, Cao Y, Ao G, Hu L, Liu H, Wu J, Wang X, Jin M, Zheng S, Zhen X, Alkayed Nj, Jia J, Cheng J. Antioxid Redox Signal. 2014 Oct 20;21(12):1741-58. Doi: 10.1089/Ars.2013.5587. Epub 2014 Mar 13. Camkk??-Dependent Activation Of Amp-Activated Prote...
Sun Hx, Chen Lq, Zhang J, Chen Fy. J Ethnopharmacol. 2014 May 14;153(3):831-9. Doi: 10.1016/J.Jep.2014.03.052. Epub 2014 Mar 29. Anti-Tumor And Immunomodulatory Activity Of Peptide Fraction From The Larvae Of Musca Domestica.
Ruan Z, Liu S, Zhou Y, Mi S, Liu G, Wu X, Yao K, Assaad H, Deng Z, Hou Y, Wu G, Yin Y. Plos One. 2014 Jun 2;9(6):E97815. Doi: 10.1371/Journal.Pone.0097815. Ecollection 2014. Chlorogenic Acid Decreases Intestinal Permeability And Increases Expressi...
Meng A, Wang B, Zhang X, Qi N, Liu D, Wu J. Inflammation. 2015;38(3):1260-6. Doi: 10.1007/S10753-014-0093-X. Additive Suppression Of Lps-Induced Il-10 And Tnf-?? By Pre-Treatment Of Dexamethasone And Sb203580 In A Murine Alveolar Macrophage Cell Li...
Chen W, Guo Y, Yang W, Zheng P, Zeng J, Tong W. Exp Brain Res. 2015 Oct;233(10):2823-31. Doi: 10.1007/S00221-015-4352-3. Epub 2015 Jun 13. Protective Effect Of Ginsenoside Rb1 On Integrity Of Blood-Brain Barrier Following Cerebral Ischemia.
Li Y, Li Sy, Li Js, Deng L, Tian Yg, Jiang Sl, Wang Y, Wang Yy. Biol Pharm Bull. 2012;35(10):1752-60. Epub 2012 Aug 3. A Rat Model For Stable Chronic Obstructive Pulmonary Disease Induced By Cigarette Smoke Inhalation And Repetitive Bacterial Infe...
Niu Z, Ma J, Chu H, Zhao Y, Feng W, Cheng Y. Neurosci Lett. 2012 Nov 7;529(2):112-7. Doi: 10.1016/J.Neulet.2012.09.034. Epub 2012 Sep 27. Melanocortin 4 Receptor Antagonists Attenuates Morphine Antinociceptive Tolerance, Astroglial Activation And ...
Liu W, Dong Xs, Sun Yq, Liu Z. Int J Clin Exp Med. 2014 Apr 15;7(4):919-26. Ecollection 2014. A Novel Fluid Resuscitation Protocol: Provide More Protection On Acute Kidney Injury During Septic Shock In Rats.
Zhang Gb, Tian Lq, Li Ym, Liao Yf, Li J, Bing Fh. Phytomedicine. 2013 Aug 15;20(11):964-8. Doi: 10.1016/J.Phymed.2013.04.009. Epub 2013 Jun 6. Protective Effect Of Homonojirimycin From Commelina Communis (Dayflower) On Influenza Virus Infection In...
Yin Y, Qin T, Yu Q, Yang Q. Amino Acids. 2014 Jul;46(7):1763-74. Doi: 10.1007/S00726-014-1735-X. Epub 2014 Apr 8. Bursopentin (Bp5) From Chicken Bursa Of Fabricius Attenuates The Immune Function Of Dendritic Cells.
Tang D, Gao J, Wang S, Yuan Z, Ye N, Chong Y, Xu C, Jiang X, Li B, Yin W, Miao Y, Wang D, Jiang K. Tumour Biol. 2015 Jul;36(7):5617-26. Doi: 10.1007/S13277-015-3233-5. Epub 2015 Mar 1. Apoptosis And Anergy Of T Cell Induced By Pancreatic Stellate ...
Chen X, Zhao C, Li X, Wang T, Li Y, Cao C, Ding Y, Dong M, Finci L, Wang Jh, Li X, Liu L. Nat Chem Biol. 2015 Jan;11(1):19-25. Doi: 10.1038/Nchembio.1657. Epub 2014 Nov 10. Terazosin Activates Pgk1 And Hsp90 To Promote Stress Resistance.
Liu R, An L, Liu G, Li X, Tang W, Chen X. Antiviral Res. 2015 Aug;120:101-11. Doi: 10.1016/J.Antiviral.2015.05.008. Epub 2015 May 27. Mouse Lung Slices: An Ex Vivo Model For The Evaluation Of Antiviral And Anti-Inflammatory Agents Against Influenz...
Dai M, Wu L, He Z, Zhang S, Chen C, Xu X, Wang P, Gruzdev A, Zeldin Dc, Wang Dw. J Cell Physiol. 2015 Sep;230(9):2108-19. Doi: 10.1002/Jcp.24939. Epoxyeicosatrienoic Acids Regulate Macrophage Polarization And Prevent Lps-Induced Cardiac Dysfunction.
Zhang Y, Zhang S, Li H, Huang M, Xu W, Chu K, Chen L, Chen X. Mol Med Rep. 2015 Jul;12(1):988-94. Doi: 10.3892/Mmr.2015.3515. Epub 2015 Mar 19. Ameliorative Effects Of Gualou Guizhi Decoction On Inflammation In Focal Cerebral Ischemic-Reperfusion ...
Shi Jq, Chen J, Wang Br, Zhu Yw, Xu Y, Wang J, Xiao H, Shi Jp, Zhang Yd, Xu J. Peptides. 2011 Aug;32(8):1617-25. Doi: 10.1016/J.Peptides.2011.06.013. Epub 2011 Jun 21. Short Amyloid-Beta Immunogens Show Strong Immunogenicity And Avoid Stimulating ...
Zhang H, Xia X, Han F, Jiang Q, Rong Y, Song D, Wang Y. Mol Pharm. 2015 May 4;12(5):1648-61. Doi: 10.1021/Acs.Molpharmaceut.5B00069. Epub 2015 Apr 2. Cathelicidin-Bf, A Novel Antimicrobial Peptide From Bungarus Fasciatus, Attenuates Disease In A D...
Wang Sb, Deng Yq, Ren J, Xiao Bk, Chen Z, Tao Zz. Scand J Immunol. 2013 Dec;78(6):507-15. Doi: 10.1111/Sji.12118. Lactoferrin Administration Into The Nostril Alleviates Murine Allergic Rhinitis And Its Mechanisms.
Ozkara S, Keles E, Ilhan N, Gungor H, Kaygusuz I, Alpay Hc. Eur Arch Otorhinolaryngol. 2012 Dec;269(12):2519-24. Doi: 10.1007/S00405-012-1967-X. Epub 2012 Feb 23. The Relationship Between Th1/Th2 Balance And 1??,25-Dihydroxyvitamin D
Berni Canani R, Paparo L, Nocerino R, Cosenza L, Pezzella V, Di Costanzo M, Capasso M, Del Monaco V, D'Argenio V, Greco L, Salvatore F. Clin Epigenetics. 2015 Mar 31;7(1):38. Doi: 10.1186/S13148-015-0070-8. Ecollection 2015. Differences In Dna Met...
Huang L, Cao J. Oxid Med Cell Longev. 2014;2014:248786. Doi: 10.1155/2014/248786. Epub 2014 Mar 5. The Protective Effects Of Shen-Fu Injection On Experimental Acute Pancreatitis In A Rat Model.
Ou J, Shi W, Xu Y, Tao Z. Ann Allergy Asthma Immunol. 2014 Dec;113(6):658-665.E1. Doi: 10.1016/J.Anai.2014.08.015. Epub 2014 Sep 17. Intranasal Immunization With Dna Vaccine Coexpressing Der P 1 And Ubiquitin In An Allergic Rhinitis Mouse Model.
Jin Q, Cheng J, Liu Y, Wu J, Wang X, Wei S, Zhou X, Qin Z, Jia J, Zhen X. Brain Behav Immun. 2014 Aug;40:131-42. Doi: 10.1016/J.Bbi.2014.03.003. Epub 2014 Mar 12. Improvement Of Functional Recovery By Chronic Metformin Treatment Is Associated With...
Ozkan Zs, Deveci D, Kumbak B, Simsek M, Ilhan F, Sekercioglu S, Sapmaz E. J Reprod Immunol. 2014 Jun;103:53-8. Doi: 10.1016/J.Jri.2013.11.002. Epub 2013 Dec 8. What Is The Impact Of Th1/Th2 Ratio, Socs3, Il17, And Il35 Levels In Unexplained Infert...
Presti I, D'Orazio G, Labra M, La Ferla B, Mezzasalma V, Bizzaro G, Giardina S, Michelotti A, Tursi F, Vassallo M, Di Gennaro P. Appl Microbiol Biotechnol. 2015 Jul;99(13):5613-26. Doi: 10.1007/S00253-015-6482-8. Epub 2015 Mar 7. Evaluation Of The...
Chen F, Ni Y, Ye Y, Sun H, Li X, Xu S. Int Immunopharmacol. 2010 Oct;10(10):1153-60. Doi: 10.1016/J.Intimp.2010.06.021. Epub 2010 Jul 4. Comparison Of Immunosuppressive Activity Of Stephanoside E And Its Aglycone From Stephanotis Mucronata In Vitro.
Yu Zm, Liu Zh, Chen J, Zeng Q. Chin J Integr Med. 2014 Dec;20(12):934-43. Doi: 10.1007/S11655-014-1863-X. Epub 2014 Nov 27. Anti-Inflammatory Effect Of Qingwen Baidu Decoction (????????????
Zhu Hc, Qiu T, Liu Xh, Dong Wc, Weng Xd, Hu Ch, Kuang Yl, Gao Rh, Dan C, Tao T. Cell Immunol. 2012;274(1-2):12-8. Doi: 10.1016/J.Cellimm.2012.02.012. Epub 2012 Mar 14. Tolerogenic Dendritic Cells Generated By Relb Silencing Using Shrna Prevent Acu...
Guida F, Luongo L, Aviello G, Palazzo E, De Chiaro M, Gatta L, Boccella S, Marabese I, Zjawiony Jk, Capasso R, Izzo Aa, De Novellis V, Maione S. Mol Pain. 2012 Aug 23;8:60. Doi: 10.1186/1744-8069-8-60. Salvinorin A Reduces Mechanical Allodynia And...
Sofian M, Aghakhani A, Farazi Aa, Banifazl M, Eslamifar A, Rashidi N, Khadem Sadegh A, Ramezani A. Hepat Mon. 2012 Dec;12(12):E6156. Doi: 10.5812/Hepatmon.6156. Epub 2012 Dec 29. Serum Profile Of T Helper 1 And T Helper 2 Cytokines In Hepatitis C ...
Customer Q&As
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.
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Keywords: component, ingredient