IL-6 ELISA Kit (Human) (OKBB00189)

Catalog#: OKBB00189

• Size: 96 Tests

• Application: ELISA
• Gene Symbol: IL-6
• Official Gene Full Name: interleukin 6
• NCBI Gene Id: 3569
• Alias Symbols: Interleukin-6, Interleukin 6, IL 6, IFNB2
• Description of Target: The human interferon-beta 2 gene (IFNB2) product is identical to that for the B-cell stimulation factor-2 (BSF-2), the hybridoma growth factor (HGF) ("interleukin-6"), and the hepatocyte stimulating factor (HSF). Proteins derived from this gene mediate the plasma protein response to tissue injury (acute-phase response) and regulate the growth and differentiation of both B and T cells.1 Interleukin-6 (IL6) has come to be regarded as a potential osteoporotic factor because it has stimulatory effects on cells of the osteoclast lineage, and, thus, may play a role in the pathogenesis of bone loss associated with estrogen deficiency.2 IL-6 has many roles essential to the regulation of the immune response, hematopoiesis, and bone resorption.3 It is involved not only in the hepatic acute phase response but also in adipose tissue metabolism, lipoprotein lipase activity, and hepatic triglyceride secretion.4 Overproduction of IL-6, a proinflammatory cytokine, is associated with a spectrum of age-related conditions including cardiovascular disease, osteoporosis, arthritis, type 2 diabetes, certain cancers, periodontal disease, frailty, and functional decline.5 BSF-2 is a novel interleukin consisting of 184 amino acids.6 The standard product used in this kit is recombinant human IL-6, consisting of 184 amino acids with the molecular mass of 20.3kDa.
• Protein Name: Interleukin-6
• Swissprot Id: P05231
• Protein Accession #: NP_000591.1
• Nucleotide Accession #: NM_000600.4
• Sample Type: cell culture supernates, serum and plasma (heparin, EDTA, citrate)
• Sensitivity: < 0.3 pg/ml
• Kit Range: 4.69 pg/ml-300 pg/ml
• Kit Reproducibility:

  	Intra-Assay Precision			Inter-Assay Precision
  Sample	1	2	3	1	2	3
  Mean (pg/mL)	16.3	98	179	18.2	99	185
  St. Dev.	0.8	2.3	4.2	1	3.6	5.7
  %CV	4.90	2.34	2.34	5.49	3.63	3.08

• Kit Duration: ~ 3 Hours
• Kit Principle: Aviva Systems Biology IL-6 ELISA Kit (Human) (OKBB00189) is based on standard sandwich enzyme-linked immune-sorbent assay technology. An antibody specific for IL-6 has been pre-coated onto 96-wellplate (12 x 8 Well Strips). Standards (E.coli, P29-M212) and test samples are added to the wells, incubated and removed. A biotinylated detector antibody specific for IL-6 is added, incubated and followed by washing. Avidin-Biotin-Peroxidase Complex is then added, incubated and unbound conjugate is washed away. An enzymatic reaction is visualized through the addition of TMB substrate which is catalyzed by HRP to produce a blue color product that changes yellow after adding acidic stop solution. The density of yellow coloration read by absorbance at 450 nm and is quantitatively proportional to the amount of sample Human IL-6 captured in well.
• Kit Component:

  Component	Amount
  Lyophilized recombinant human IL-5 standard	10 ng/tube x 2
  Anti-human IL-5 Antibody Well Plate	96 Wells
  Sample diluent buffer	30 mL
  Biotinylated anti- human IL-5 antibody	130 uL, dilution 1:100
  Antibody diluent buffer	12 mL
  Avidin-Biotin-Peroxidase Complex (ABC)	130 uL, dilution 1:100
  ABC diluent buffer	12 mL
  TMB color developing agent	10 mL
  TMB stop solution	10 mL

• Kit Detection Method: Colorimetric, OD450 nm
• RNA Seq: Find tissues and cell lines supported by RNA-seq analysis to express IL-6.
• Predicted Homology Based on Immunogen Sequence: No detectable cross-reactivity with any other cytokine.
• Datasheets/Manuals: Click here to download product manual. As variation between lots may occur, always reference the lot-specific manual received with each kit.
• Reconstitution and Storage: Store as indicated in product manual.
• Target Reference: 1. Bowcock, A. M.; Kidd, J. R.; Lathrop, G. M.; Daneshvar, L.; May, L. T.; Ray, A.; Sehgal, P. B.; Kidd, K. K.; Cavalli-Sforza, L. L. The human
  'interferon-beta-2/hepatocyte stimulating factor/interleukin-6' gene: DNA polymorphism studies and localization to chromosome 7p21.
  Genomics 3: 8-16, 1988.
  2. Ota, N.; Nakajima, T.; Nakazawa, I.; Suzuki, T.; Hosoi, T.; Orimo, H.; Inoue, S.; Shirai, Y.; Emi, M. A nucleotide variant in the promoter region
  of the interleukin-6 gene associated with decreased bone mineral density. J. Hum. Genet. 46: 267-272, 2001. 3. Chung, H. W.; Seo, J.-S.; Hur, S. E.; Kim, H. L.; Kim, J. Y.; Jung, J. H.; Kim, L. H.; Park, B. L.; Shin, H. D. Association of interleukin-6
  promoter variant with bone mineral density in pre-menopausal women. J. Hum. Genet. 48: 243-248, 2003.
  4. Ferrari, S. L.; Ahn-Luong, L.; Garnero, P.; Humphries, S. E.; Greenspan, S. L. Two promoter polymorphisms regulating interleukin-6 gene
  expression are associated with circulating levels of C-reactive protein and markers of bone resorption in postmenopausal women. J. Clin.
  Endocr. Metab. 88: 255-259, 2003.
  5. Kiecolt-Glaser, J. K.; Preacher, K. J.; MacCullum, R. C.; Atkinson, C.; Malarkey, W. B.; Glaser, R. Chronic stress and age-related increases
  in the proinflammatory cytokine IL-6. Proc. Nat. Acad. Sci. 100: 9090-9095, 2003.
  6. Hirano, T.; Yasukawa, K.; Harada, H.; Taga, T.; Watanabe, Y.; Matsuda, T.; Kashiwamura, S.; Nakajima, K.; Koyama, K.; Iwamatsu, A.;
  Tsunasawa, S.; Sakiyama, F.; Matsui, H.; Takahara, Y.; Taniguchi, T.; Kishimoto, T. Complementary DNA for a novel human interleukin
  (BSF-2) that induces B lymphocytes to produce immunoglobulin. Nature 324: 73-76, 1986.
• Specificity: There is no detectable cross-reactivity with other relevant proteins.
• Publications:

  Selfe, J. et al. The effect of three different (-135C) whole body cryotherapy exposure durations on elite rugby league players. PLoS One 9, e86420 (2014). ELISA, Human 24489726

  Ota, H. et al. Pathological mechanism of musculoskeletal manifestations associated with CRPS type II: An animal study. Pain (2014). doi:10.1016/j.pain.2014.06.016 ELISA, Human 25016218

• Additional Information: Range: 4.69pg/ml-300pg/ml
  Principle: Aviva’s human IL-6 ELISA Kit was based on standard sandwich
  enzyme-linked immune-sorbent assay technology. Human IL-6 specific-specific
  monoclonal antibodies were precoated onto 96-well plates. The human specific
  detection polyclonal antibodies were biotinylated. The test samples and
  biotinylated detection antibodies were added to the wells subsequently and
  then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase
  Complex was added and unbound conjugates were washed away with PBS or
  TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction.
  TMB was catalyzed by HRP to produce a blue color product that changed into
  yellow after adding acidic stop solution. The density of yellow is proportional to
  the human IL-6 amount of sample captured in plate
  Notes: 1. To inspect the validity of experiment operation and the appropriateness of sample dilution proportion, pilot
  experiment using standards and a small number of samples is recommended.
  2. The TMB Color Developing agent is colorless and transparent before using, contact us freely if it is not the case.
  3. Before using the Kit, spin tubes and bring down all components to the bottom of tubes.
  4. Duplicate well assay is recommended for both standard and sample testing.
  5. Don’t let 96-well plate dry, for dry plate will inactivate active components on plate.
  6. Don’t reuse tips and tubes to avoid cross contamination.
  7. To avoid to use the reagents from different batches together.
  8. In order to avoid marginal effect of plate incubation due to temperature difference ( reaction may be stronger in the
  marginal wells), it is suggested that the diluted ABC and TMB solution will be pre-warmed in 37C for 30 min before
  using.
  Background: The human interferon-beta 2 gene (IFNB2) product is identical to that for the B-cell stimulation factor-2 (BSF-2), the hybridoma growth factor (HGF) ("interleukin-6"), and the hepatocyte stimulating factor (HSF). Proteins derived from this gene mediate the plasma protein response to tissue injury (acute-phase response) and regulate the growth and differentiation of both B and T cells.1 Interleukin-6 (IL6) has come to be regarded as a potential osteoporotic factor because it has stimulatory effects on cells of the osteoclast lineage, and, thus, may play a role in the pathogenesis of bone loss associated with estrogen deficiency.2 IL-6 has many roles essential to the regulation of the immune response, hematopoiesis, and bone resorption.3 It is involved not only in the hepatic acute phase response but also in adipose tissue metabolism, lipoprotein lipase activity, and hepatic triglyceride secretion.4 Overproduction of IL-6, a proinflammatory cytokine, is associated with a spectrum of age-related conditions including cardiovascular disease, osteoporosis, arthritis, type 2 diabetes, certain cancers, periodontal disease, frailty, and functional decline.5 BSF-2 is a novel interleukin consisting of 184 amino acids.6 The standard product used in this kit is recombinant human IL-6, consisting of 184 amino acids with the molecular mass of 20.3kDa.

Protocol:

Tips Information: