Rat Interleukin 6,IL-6 ELISA KIT

Instructions
Code CSB-E04640r
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

This Rat Interleukin 6 (IL-6) ELISA Kit is suitable for qualitatively determining IL-6 concentrations in muliple biological fluids, including rat serum, plasma, and tissue homogenates in vitro. IL-6 is a pleiotropic pro-inflammatory cytokine, and its production is responsive to tissue injury and infections. It is involved in diverse biological processes such as the modulation of the host immune response, the progression of several virus infectons, and the regulation of immune cell proliferation and differentiation. IL-6 exerts its biological roles through a hexameric complex IL-6/IL-6R/gp130, which activates different signaling pathways such as JAK/STAT3, Ras/MAPK, PI3K–PKB/Akt to execute various biochemical functions. Deregulation of IL-6 signaling is associated with chronic inflammation, and multifactorial auto-immune disorders.

This kit uses the quantitative sandwich-based enzyme immunoassay technique to measure the amount of rat IL-6 in the sample. Standards and samples are respectively added to the microplate wells pre-coated with anti-rat IL-6 antibody. Biotin-labeled IL-6 antibody, HRP-avidin, TMB substrate are pipped into the microplate in turn. The capture antibody pre-coated on the plate captures the IL-6 in the rat samples. IL-6 binds to the biotinylated anti-rat IL-6 monoclonal antibody. And the biotin on the biotinylated anti-rat IL-6 monoclonal antibody binds to the avidin on the enzyme label, forming immune complexes. The color renders blue after the addition of TMB substrate. The addition of stop solution into the wells immediately turns the blue into yellow. The concentration of IL-6 in the samples is directly proportional to OD (450nm). Each manufactured lot of this ELISA kit was quality tested for criteria such as sensitivity, specificity, and lot-to-lot consistency. See manual for more information on validation.

Alternative Names Il6 ELISA Kit; Il-6 ELISA Kit; Interleukin-6 ELISA Kit; IL-6 ELISA Kit
Abbreviation IL6
Uniprot No. P20607
Species Rattus norvegicus (Rat)
Sample Types serum, plasma, tissue homogenates
Detection Range 0.312 pg/mL-20 pg/mL
Sensitivity 0.078 pg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Immunology
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10%
Three samples of known concentration were tested in twenty assays to assess.
Linearity

To assess the linearity of the assay, samples were spiked with high concentrations of rat IL-6 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.

  Sample Serum(n=4)
1:2 Average % 86
  Range % 84-95
1:4 Average % 95
  Range % 91-100
1:8 Average % 84
  Range % 81-89
1:16 Average % 92
  Range % 87-96
Recovery

The recovery of rat IL-6 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
 

Sample Type Average % Recovery Range
Serum (n=5)   95 89-100
EDTA plasma (n=4) 96 92-99
Typical Data

These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.

pg/ml OD1 OD2 Average Corrected
20 2.524 2.615 2.570 2.420
10 2.273 2.247 2.260 2.110
5 1.762 1.845 1.804 1.654
2.5 1.156 1.214 1.185 1.035
1.25 0.687 0.674 0.681 0.531
0.625 0.356 0.369 0.363 0.213
0.312 0.258 0.239 0.249 0.099
0 0.147 0.152 0.150  
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with anti-rat IL-6 antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard --- Dilute a bottle of standard at dilution series, read the OD values, and then draw a standard curve.
  • Biotin-labeled IL-6 antibody (100 x concentrate) 1 x 120 μl ---Act as the detector antibody.
  • HRP-avidin (100 x concentrate) 1 x 120 μl --- Avidin binds to the detection antibody, and HRP catalyzes the TMB substrate and thus changes the solution color.
  • Biotin-antibody Diluent 1 x 15 ml ---Dilute the Biotin-antibody solution.
  • HRP-avidin Diluent 1 x 15 ml ---Dilute the HRP-avidin solution.
  • Sample Diluent 1 x 50 ml---Dilute the sample to an appropriate concentration.
  • Wash Buffer (25 x concentrate) 1 x 20 ml --- Wash away unbound or free substances.
  • TMB Substrate 1 x 10 ml --- The TMB-HRP coproduct appears blue after adding the chromogenic agent. After the enzyme reaction is terminated, the TMB product changes from blue to yellow. It can be quantified in the colorimeter.
  • Stop Solution 1 x 10 ml --- Stop the color reaction.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

Citations

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 Q&A
Q:

Seeking quotation for ELISA kits of IL-6. I am working on SD Rats and project related to High Fat Diet induced Metabolic complications.

A:
Thanks for your inquiry.
We have this kit available:
#CSB-E04640r,Rat Interleukin 6,IL-6 ELISA KIT
Usually this kit is used to test commmon rats, but it can also detect SD Rats.Pls follow your actual test result. I will send you the kit manual by email and ask the local distributor to give you a quote soon.

Target Data

Function Cytokine with a wide variety of biological functions. It is a potent inducer of the acute phase response. Plays an essential role in the final differentiation of B-cells into Ig-secreting cells Involved in lymphocyte and monocyte differentiation. Acts on B-cells, T-cells, hepatocytes, hematopoietic progenitor cells and cells of the CNS. Required for the generation of T(H)17 cells. Also acts as a myokine. It is discharged into the bloodstream after muscle contraction and acts to increase the breakdown of fats and to improve insulin resistance. It induces myeloma and plasmacytoma growth and induces nerve cells differentiation (By similarity).
Gene References into Functions
  1. In summary, this study demonstrated that IL-6 enhances mitochondrial biogenesis and survival in astrocytes under experimental septic conditions. Additionally, IL-6 plays the pro-survival role through IL-6/AMPK signaling pathway. PMID: 29674256
  2. Aerobic exercise improves the lipid metabolism disorder and interferes with germ cell apoptosis by reducing interleukin-6 and interleukin-6 receptor expression in nutritional obesity rat model. PMID: 28776758
  3. These findings indicated that IL-6 might exert dual effects on tendon-derived stem cells in vitro: strongly enhancing their proliferation but inhibiting their tenogenic differentiation via the JAK/Stat3 pathway. PMID: 29547593
  4. in adjuvant arthritis fibroblast-like synoviocytes, MeCP2 mediates hypermethylation of PTCH1 gene and decreases the expression of PTCH1 protein, thus activating Hedgehog signaling pathway and increasing secretion of IL-6 and TNF-alpha. PMID: 28573530
  5. IL-6 expression increased in all groups compared to control. Lgr5 expression was up-regulated in cells exposed to a single stressor, but down-regulated when multiple stressors were administered. PMID: 29018960
  6. IL-6 synthesis and secretion were connected to acute peritoneal dialysis fluid exposure, and this response was triggered by TRPA1 receptors, possibly located to non-neuronal cells. PMID: 28698251
  7. The interleukin 6 (IL-6)-174 polymorphism was significantly associated with prostate cancer in Slovak patients. PMID: 29848725
  8. The present study demonstrated that IL-6 RNA interference may protect the lung from acute lung injury induced by intestinal ischemia/reperfusion, and that this protective role may be associated with upregulation of IL-10. PMID: 28713893
  9. In the present study, isolated rat peritoneal mesothelial cells were treated with TGF-beta1 and/or IL-6 which elicited mesenchymal transition (MMT) as determined by morphologic and biochemical techniques ..Partial characterization of relevant signaling pathways identified Erk1/2 activation, which was enhanced by combined TGF-b1/IL-6 administration, as a crucial necessary factor in the transition.. PMID: 26825317
  10. Comparative Modulation of Levels of Oxidative Stress in the Liver of Anti-Tuberculosis Drug Treated Wistar Rats by Vitamin B12, Beta-Carotene, and Spirulina fusiformis: Role of NF-kappaB, iNOS, IL-6, and IL-10.( PMID: 28387444
  11. An intracellular adrenomedullin system reduces IL-6 release via a NF-kB-mediated, cAMP-independent transcriptional mechanism in rat thymic epithelial cells. PMID: 27619517
  12. Crocin attenuate Tumor Necrosis Factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in streptozotocin-induced diabetic rat aorta/ PMID: 27529541
  13. Administration of Methylglyoxal led to depletion of antioxidant enzymes, induction of fibrosis (p < 0.001), up-regulated expression of RAGE (3.5 fold), TGF-beta (4.4 fold), SMAD2 (3.7 fold), SMAD3 (6.0 fold), IL-6 (4.3 fold) and TNF-alpha (5.5 fold) in the heart tissues compared to control rats. PMID: 28091942
  14. After being treated with calcitriol following hemorrhagic shock resulted in better survival rate, lower serum levels of TNF-alpha and IL-6, and lesser hepatic, renal, and pulmonary histo-pathologic scores of injury in rats. PMID: 27180201
  15. TNF-alpha, IL-6 and IL-10 expressions, responsible for disparity in action of curcumin against cisplatin-induced nephrotoxicity in rats. PMID: 28258441
  16. there may be dysregulation in IL-6 production from astrocytes. PMID: 27539920
  17. Local cryotherapy's local and systemic anti-inflammatory effects are mainly IL-6/IL-17A-driven and TNF-alpha independent in the adjuvant-induced arthritis model PMID: 28759646
  18. This study suggests direct stimulation of autophagy as a novel mechanism for IL-6-mediated protection of beta cells from stress-induced apoptosis. PMID: 28592636
  19. These data support prior findings suggesting that IL-6 mediates the leptin-sensitizing effects of amylin on VMN neurons and that the inherent leptin resistance of DIO rats can be effectively reversed at a cellular level by IL-6. PMID: 27534878
  20. Exercise combined with ultrasound attenuates neuropathic pain in rats and is associated with downregulation of IL-6 and TNF-alpha, and with upregulation of IL-10. PMID: 28319508
  21. Two-way ANOVA showed an interaction of prenatal low protein and postnatal high fat energy diet on IL-6 and IL-1beta transcription. PMID: 28141871
  22. Study provided preliminary evidence showing that high frequency vagal stimulation attenuates the expression of early onset pro-inflammatory cytokines IL-6 and TNFalpha in a neonatal rat model of airway infection. PMID: 27049312
  23. Overexpression of IL-6 is sufficient to induce myofibroblastic proliferation, differentiation, and fibrosis, probably via increased TGF-beta1-mediated MMP2/MMP3 signaling during ischemic myocardial remodeling monoculture or coculture models under hypoxic conditions. PMID: 27348628
  24. this paper shows that antidepressant rhodioloside decreased pro-inflammatory cytokine interleukin-6 PMID: 27214337
  25. Data show that cardiac arrest (CA) increased HIF-1alpha and pro-inflammatory cytokines (PICs) including IL-6 and TNF-alpha in the hippocampus CA1 region. PMID: 27383646
  26. the release of IL6 from MSCs was regulated via the TLR2/NFkappaB signalling pathway. Endogenous IL6 reduced apoptosis and protected OGDinjured PC12 cells when they were cocultured with MSCs. PMID: 27108485
  27. The levels of TNF-alpha and IL-6 and the intestinal wet/dry weight ratios were higher in the subarachnoid hemorrhage groups compared to the control group. PMID: 27993875
  28. low-level laser therapy (LLLT) administered following muscle injury modulates the mRNA expression of MyoD and myogenin. Moreover, the both forms of LLLT administration were able to modulate the mRNA expression of IL-6 during the muscle repair process. PMID: 26914683
  29. our results indicate that UC-MSCs can alleviate insulin resistance in part via production of IL-6 that elicits M2 polarization. Additionally, human obesity and insulin resistance were associated with increased pro-inflammatory ATMs infiltration. Thus, MSCs may be a new treatment for obesity-related insulin resistance and T2D concerning macrophage polarized effects PMID: 26523620
  30. Results suggest that activated-p38[alpha]-regulated IL-6 synthesis constitutes a neuroprotective mechanism of NaHS against oxygen-glucose deprivation/reoxygenation-induced neuronal injury in PC12 cells PMID: 26760196
  31. decrease in all groups after manipulation. Data show that the postoperative interleukin 6 (IL-6) levels were significantly increased in the conventional cecum resection (CCR) group compared to the laparoscopic cecum resection (LCR) group. PMID: 26846568
  32. Ang III, similar to Ang II, has pro-inflammatory effects since it induces STAT3 leading to an induction of IL-6 mRNA expression. PMID: 24961501
  33. the adenylyl cyclase/cAMP/PKA pathway upregulates IL-1beta-induced IL-6 synthesis through enhancement of the JAK2/STAT3 pathway in C6 glioma cells PMID: 26527061
  34. LPS significantly increased the supernatant TNF-alpha and IL-6 concentrations and reduced TH protein expression in PC12 cells, while silencing Nogo-A could block these effects PMID: 26583134
  35. Data show that expressions of vascular endothelial growth factor, interleukin 6, but not toll-like receptor 4 were modulated in everolimus-treated group. PMID: 26390417
  36. Data show that the ratio of peripheral blood CD4(+)/CD8(+) T lymphocytes and the levels of connexin-40 (Cx40), interleukins IL-2, IL-4 and IL-6 are significantly elevated in SH rats. PMID: 26927369
  37. Data show pioglitazone inhibits inflammatory reaction by up-regulating the level of peroxisome proliferator activated receptor gamma (PPARgamma) mRNA and down-regulating the levels of inflammatory cytokines TNF-alpha and IL-6 mRNAs in traumatic brain injury (TBI). PMID: 26927377
  38. Diabetic rats show an increase in the contractions of distal colon partly via the IL-6-IL-6 receptor pathway. PMID: 26191141
  39. REV-ERBalpha has a role in circadian oscillation in the expressions of Alas1, Ppargc1a, and Il6 genes in rat granulosa cells PMID: 26102301
  40. Data show that the negative regulation of cardiac fibroblasts (CFs) differentiation by qiliqiangxin (QL) through an interleukin-6(IL-6) transcriptional mechanism. PMID: 25752645
  41. IL-6 activates PI3K and PKCzeta signaling and determines cardiac differentiation in rat embryonic H9c2 cells PMID: 26205888
  42. Knockdown of IL-1Beta increased IL-6 expression in cerebral hypoxia-ischemia. PMID: 24965599
  43. Early increase of calpain-2 in L5-ventral root transection rats might be responsible for the induction of allodynia via up-regulating IL-6 in DRG neurons. PMID: 25150005
  44. HGF differentially regulates IL-6 production in hepatocytes (induces) versus immune cells (suppresses), signifying disparate regulation of the cell sources involved in IL-6 production is a biologically relevant mechanism. PMID: 24763697
  45. Neuromodulatory effects of interleukin-6 (IL-6) and corticotropin-releasing factor receptor (CRFR) 1 in visceral pain and stress-induced defecation in the WKY rat model of irritable bowel syndrome were investigated. PMID: 25260633
  46. Studied Sporothrix schenckii induction of release of tumor necrosis factor alpha (TNF-alpha) and interleukin (IL)-6 and activation of the extracellular signal-regulated kinase (ERK) signaling pathway in mast cells. PMID: 25262023
  47. S100beta-positive dendritic-like cells can sense an increase in extracellular protons via GPR68 and respond by the production of IL-6 in order to suppress the up-regulation of Pomc expression. PMID: 25129106
  48. Interleukin 6 (IL-6) levels rapidly increased within 24 h after SAH, reached a peak 24 h after SAH, and decreased slightly at 48 h. SOCS-3 was involved in cerebral vasospasm after subarachnoid hemorrhage via an IL-6- and IL-10-related mechanism. PMID: 24463741
  49. The neuronal apoptosis rate was positively correlated with the levels of TNF-alpha and IL-6, and negatively correlated with IL-10 and GDNF. PMID: 24706318
  50. The continuing increase in the production of IL-6 in culture was accompanied by a decreased formation of erythroblastic islets de novo. PMID: 25682689

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Subcellular Location Secreted
Protein Families IL-6 superfamily
Database Links

KEGG: rno:24498

STRING: 10116.ENSRNOP00000013732

UniGene: Rn.9873

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