Human IL-6 ELISA Kit (Colorimetric)

Product Discontinued

Human IL-6 ELISA Kit (Colorimetric) Summary

• Description: IL-6 ELISA Kit (NBP2-31051) is provided as 2 x 96 well format and it is good for 192 tests. This kit measures free IL-6 in the supernatant of activated cells and its detection is based on sandwich ELISA principle. An anti-IL-6 monoclonal antibody is coated on a plate which then captures free IL-6. The bound IL-6 is detected by adding a second, biotinylated, monoclonal IL-6 antibody followed by HRP-conjugated streptavidin. ABTS substrate is then added and the concentration is determined by colorimetric detection in an ELISA plate reader.
• Specificity: Specific for free human IL6 in cell supernatant of activated cells.
• Standard Curve Range: 15.6-1000 pg/ml
• Sensitivity: 2 pg/ml
• Assay Type: Sandwich ELISA
• Sample Volume: 0.1 ml
• Kit Type: ELISA Kit (Colorimetric)
• Gene: IL6

Applications/Dilutions

• Dilutions:
  • ELISA
• Application Notes: Human IL-6 ActivELISA For the detection of human IL-6. The Human IL-6 ActivELISA(TM) Kit measures free IL-6 in the supernatant of activated cells. Standard protocols for detecting IL-6 activity include ELISA and Western Blot. The IL-6 ActivELISA(TM) can be completed in one day using a sandwich ELISA protocol. An anti-IL-6 monoclonal antibody is coated on a plate which then captures free IL-6. The bound IL-6 is detected by adding a second, biotinylated, monoclonal IL-6 antibody followed by HRP-conjugated streptavidin. ABTS substrate is then added and the concentration is determined by colorimetric detection in an ELISA plate reader.

Packaging, Storage & Formulations

• Storage: Storage is content dependent.

Notes

Additional items required for the ELISA (not included in the IL-6 ActivELISA Module): 96-well ELISA plate reader

Alternate Names for Human IL-6 ELISA Kit (Colorimetric)

• B-cell differentiation factor
• B-cell stimulatory factor 2
• BSF2
• BSF-2
• CDF
• CTL differentiation factor
• HSF
• hybridoma growth factor
• IFNB2
• IFN-beta-2
• IL6
• IL-6
• Interferon beta-2
• interleukin 6 (interferon, beta 2)
• interleukin BSF-2
• interleukin-6
• MGI-2A

Background

IL-6 is a cytokine that functions in inflammation, and maturation of B cells. The protein is primarily produced at sites of acute and chronic inflammation, where it is secreted into the serum and induces a transcriptional inflammatory response. This classical responsiveness to IL-6 is governed by a receptor complex consisting of two membrane-bound subunits, an 80-kDa cognate Alpha-chain (IL-6R-Alpha), and a ubiquitously expressed 130-kDa Beta-chain receptor (gp130) which acts as the universal signal transducing element for all IL-6 family cytokines. Alternatively, IL-6 regulation of leukocyte trafficking relies upon signaling via its soluble IL-6R Alpha (termed IL-6 trans-signaling) (Fielding CA et al, 2008). IL-6 trans-signals via STAT3 which acts as a critical modulator of LPS-driven proinflammatory responses through cross-talk regulation of the TLR4/Mal signaling pathway, and potentially implicate cross-talk between JAK/STAT and TLR pathways as a broader mechanism that regulates the severity of the host inflammatory response (Greenhill CJ et al, 2011). After this initial wave of proinflammatory cytokine production, antiinflammatory cytokines such as TGF-Beta and IL-10 are produced to dampen the TLR4-driven production of proinflammatory cytokines. Thus functioning of this gene is implicated in a wide variety of inflammation-associated disease states, including susceptibility to diabetes mellitus and systemic juvenile rheumatoid arthritis (Takeda K et al, 2003).

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. ELISA Kits are guaranteed for 6 months from date of receipt.

Publications for IL-6 ELISA Kit (NBP2-31051)(5)

Publications using NBP2-31051

• Deleon-Pennell KY, Ero OK, Ma Y et al. Glycoproteomic Profiling Provides Candidate Myocardial Infarction Predictors of Later Progression to Heart Failure ACS Omega 2019-01-31 [PMID: 30729226] (ELISA, Human)
• Ji L, Hou X, Liu W et al. Paeoniflorin inhibits activation of the IRAK1-NF-kB signaling pathway in peritoneal macrophages from lupus-prone MRL/lpr mice Microb. Pathog. 2018-08-24 [PMID: 30149133] (ELISA, Mouse)
• Chiu Chun-Hung, Chyau Charng-Cherng, Chen Chin-Chu et al. Erinacine A-Enriched Hericium erinaceus Mycelium Produces Antidepressant-Like Effects through Modulating BDNF/PI3K/Akt/GSK-3B Signaling in Mice. International Journal of Molecular Sciences 2018-01-01 [PMID: 29364170] (ELISA, Mouse)
• Vossenkamper A, Marches O, Fairclough PD et al. Inhibition of NF-kB signaling in human dendritic cells by the enteropathogenic Escherichia coli effector protein NleE. J Immunol. 2010-10-01 [PMID: 20833837] (Human)
  
  Details:
  Nuclear and cytosolic extracts from monocyte derived human dendritic cells in the presence of EPEC LPS, Fig 3D.
• Kondo A, Yamashita T, Tamura H et al. Interferon-gamma and tumor necrosis factor-alpha induce an immunoinhibitory molecule, B7-H1, via nuclear factor-kappaB activation in blasts in myelodysplastic syndromes. Blood. 2010-08-19 [PMID: 20472834]
  
  Details:
  NF-kB/p65 ActivELISA (IMK-503) kit and Nuclear extraction kit (10081K). SKM-1 leukemic cells were separated into cytoplasmic and nuclear fractions then subject to western blot analysis, Fig 2A &B.

Review for IL-6 ELISA Kit (NBP2-31051) (1)

Average Rating: 4

(Based on 1 review)

reviewed by: Kristine DeLeon-Pennell 05/25/2017

Summary

• Sample Tested: human post-MI plasma

Comments

• Comments: Detection limit was outside the measurable range in post-MI plasma. Had to dilute standards to ensure I was able to calculate concentrations. Protocol was easy to follow.

FAQs for IL-6 ELISA Kit (NBP2-31051). (Showing 1 - 3 of 3 FAQs).

1. I was wondering if you know of any blocking antibodies against IL-6 or VEGF-C that can be used in cell culture to quench these cytokines?
   • We have no antibodies that are currently known to show blocking antibodies to VEGF-C. Please see this link to our IL-6 antibodies with blocking/neutralizing activity can be found at this link.
2. Can we use this product for serum and plasma sample? We can’t found information about usable sample type in datasheet.
   • According to the datasheet and manual of this kit, it is only recommended for assaying with the culture supernatants of the activated (cell line or primary) cells. This kit only guarantees to produce the positive results for this type of samples. However in principle, any IL-6 containing sources, like serum and plasma samples you mentioned, can also be used; except that the serum and plasma containing more proteins and other biological compounds and therefore they are more viscous than the supernatants.  This means, the diffusion of the protein target IL-6 through serum/plasma will be slower than going through the cell supernatants; and therefore the experimental conditions, such as the treating time of samples adding to the capture antibody, etc., will need to be optimized. For this part I don't have a clue. This idea can be supported by our another IL-6 kit NBP1-89869, which is applicable to the serum and plasma samples. In its datasheet (PDF), there has no pre-treatment of serum/plasma samples before applying to the microtiter plate containing the capture antibody. This suggests that there has no inhibitor or blocker of any kind to prevent the immobilized antibody from binding the freely floating protein target IL-6.
3. Over what range is IL-6 ELISA kit linear?
   • In terms of the linear region, in concentration, that can be used for measurements, Table 1 of the manual has a part of the answer: the standard curves are set up from 15.6 pg/ml to 1,000 pg/m. Then the figure in P5 has the plot of a typical standard curve, OD at 405 nm vs. the log of pg/ml. This standard curve is a liner line that could be used to determine the IL-6 concentrations in the samples, after the appropriate dilution. In short, I anticipate that this kit has been optimized to use the standard curve derived from 15.6 - 1,000 pg/ml with their respective OD readings.

Bioinformatics

• Gene Symbol: IL6