Mouse Transforming Growth factor β1,TGF-β1 ELISA kit

Code CSB-E04726m
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Mouse Transforming Growth factor β1 (TGF-β1) ELISA kit is a solid phase immunoassay specially designed to quatitatively measure mouse TGF-β1 in serum, plasma, or cell culture supernates. It is based on the Sandwich-ELISA mechainsm. TGF-β1 in the sample is bound to the capture antibody immobilized on the 96-well strip plate and then sandwiched with the biotinylated TGF-β1 antibody. After the addition of HRP-avidin and TMB substrate, the solution in the wells turns blue. The color reaction is stopped by adding the stop solution into the wells, and the color changes from blue to yellow. The color intensity is positively proportional to the TGF-β1 bound in the initial step. The TGF-β1 concentration can be calculated according to the standard curve. This kit is tested with high sensitivity, strong specificity, good linearity, precision less than 10%, recovery with 87%-100%, and lot-to-lot cosistency. More validation information is described in the product instructions.

TGF-β1 is one of TGF-β isoforms. TGF-β1 is the most common in the immune system, in which it acts as a crucial pleiotropic cytokine with potent immunoregulatory properties. It has a multitude of functions, including both promoting the formation and inhibiting the breakdown of connective tissue. It also inhibits epithelial cell proliferation. Additionally, TGF-β1 plays an important role in both normal and abnormal wound healing. TGF-β1/TGF-βRII signaling mediates a wide range of pathways, including the Erk/MAPK pathway and the PI3K/Akt pathway, finally promoting cell proliferation, differentiation, and migration.

Target Name transforming growth factor, beta 1
Alternative Names Tgfb1 ELISA Kit; Transforming growth factor beta-1 proprotein [Cleaved into: Latency-associated peptide ELISA Kit; LAP); Transforming growth factor beta-1 ELISA Kit; TGF-beta-1)] ELISA Kit
Abbreviation TGFB1
Uniprot No. P04202
Species Mus musculus (Mouse)
Sample Types serum, plasma, cell culture supernates
Detection Range 0.78 ng/mL-50 ng/mL
Sensitivity 0.2 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Signal Transduction
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 mouse TGF-β1 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)  
1:1 Average % 99  
Range % 94-104  
1:2 Average % 89  
Range % 83-97  
1:4 Average % 92  
Range % 87-97  
1:8 Average % 92  
Range % 87-96  
Recovery
The recovery of mouse TGF-β1 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) 92 87-99  
EDTA plasma (n=4) 96 90-100  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
ng/ml OD1 OD2 Average Corrected  
50 2.654 2.636 2.645 2.515  
25 2.318 2.455 2.387 2.257  
12.5 1.756 1.886 1.821 1.691  
6.25 0.997 1.001 0.999 0.869  
3.12 0.542 0.512 0.527 0.397  
1.56 0.301 0.309 0.305 0.175  
0.78 0.228 0.238 0.233 0.103  
0 0.129 0.131 0.130    
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with anti-mouse TGF-β1 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 TGF-β1 antibody (100 x concentrate) 1 x 120 μl ---Act as the detection antibody.
  • HRP-avidin (100 x concentrate) 1 x 120 μl --- Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • Biotin-antibody Diluent 1 x 15 ml ---Dilute the high concentration Biotin-antibody to an appropriate working solution.
  • HRP-avidin Diluent 1 x 15 ml---Dilute the high concentration HRP-avidin solution to an appropriate 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 --- Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • Stop Solution 1 x 10 ml --- Stop the color reaction. The solution color immediately turns from blue to yellow.
  • 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
ELISA Data Analysis Watch ELISA data processing video & download Curve Expert if needed
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

Citations

Customer Reviews and Q&A

 Customer Reviews
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Sample type: Cell culture supernatant

Sample species: Mouse

Sample dilution: No dilution

Review: I used CSB-E04726m to detect tgfβ levels in cell supernatant exosomes and mouse supernatant, respectively. The results of the standard curve were good.

By Anonymous

Target Background

Function
(From Uniprot)
Transforming growth factor beta-1 proprotein: Precursor of the Latency-associated peptide (LAP) and Transforming growth factor beta-1 (TGF-beta-1) chains, which constitute the regulatory and active subunit of TGF-beta-1, respectively.; Required to maintain the Transforming growth factor beta-1 (TGF-beta-1) chain in a latent state during storage in extracellular matrix. Associates non-covalently with TGF-beta-1 and regulates its activation via interaction with 'milieu molecules', such as LTBP1, LRRC32/GARP and LRRC33/NRROS, that control activation of TGF-beta-1. Interaction with LRRC33/NRROS regulates activation of TGF-beta-1 in macrophages and microglia. Interaction with LRRC32/GARP controls activation of TGF-beta-1 on the surface of activated regulatory T-cells (Tregs). Interaction with integrins (ITGAV:ITGB6 or ITGAV:ITGB8) results in distortion of the Latency-associated peptide chain and subsequent release of the active TGF-beta-1.; Multifunctional protein that regulates the growth and differentiation of various cell types and is involved in various processes, such as normal development, immune function, microglia function and responses to neurodegeneration. Activation into mature form follows different steps: following cleavage of the proprotein in the Golgi apparatus, Latency-associated peptide (LAP) and Transforming growth factor beta-1 (TGF-beta-1) chains remain non-covalently linked rendering TGF-beta-1 inactive during storage in extracellular matrix. At the same time, LAP chain interacts with 'milieu molecules', such as LTBP1, LRRC32/GARP and LRRC33/NRROS that control activation of TGF-beta-1 and maintain it in a latent state during storage in extracellular milieus. TGF-beta-1 is released from LAP by integrins (ITGAV:ITGB6 or ITGAV:ITGB8): integrin-binding to LAP stabilizes an alternative conformation of the LAP bowtie tail and results in distortion of the LAP chain and subsequent release of the active TGF-beta-1. Once activated following release of LAP, TGF-beta-1 acts by binding to TGF-beta receptors (TGFBR1 and TGFBR2), which transduce signal. While expressed by many cells types, TGF-beta-1 only has a very localized range of action within cell environment thanks to fine regulation of its activation by Latency-associated peptide chain (LAP) and 'milieu molecules'. Plays an important role in bone remodeling: acts as a potent stimulator of osteoblastic bone formation, causing chemotaxis, proliferation and differentiation in committed osteoblasts. Can promote either T-helper 17 cells (Th17) or regulatory T-cells (Treg) lineage differentiation in a concentration-dependent manner. At high concentrations, leads to FOXP3-mediated suppression of RORC and down-regulation of IL-17 expression, favoring Treg cell development. At low concentrations in concert with IL-6 and IL-21, leads to expression of the IL-17 and IL-23 receptors, favoring differentiation to Th17 cells. Stimulates sustained production of collagen through the activation of CREB3L1 by regulated intramembrane proteolysis (RIP). Mediates SMAD2/3 activation by inducing its phosphorylation and subsequent translocation to the nucleus. Can induce epithelial-to-mesenchymal transition (EMT) and cell migration in various cell types.
Gene References into Functions
  1. this paper shows that epithelial-derived TGF-beta1 acts as a pro-viral factor in the lung during influenza A infection PMID: 29067998
  2. Study points toward elevated levels of active TGF-beta as inducers and promoters of ectopic bone formation, and suggest that TGF-beta might be a therapeutic target in heterotopic ossification. PMID: 29416028
  3. The comparison of transforming growth factor beta family (TGFbeta) expression showed significantly higher levels of Tgfbeta3 transcript between nude and Balb/c mice but no differences were detected for Tgfbeta1. Nude DFs were specifically sensitive to the presence of the pro-regenerative TGFbeta3 isoform, showing increased collagen I deposition and alpha smooth muscle actin expression. PMID: 29637306
  4. Results indicate that the miR-23a cluster regulates osteocyte differentiation by modulating the TGF-beta signalling pathway. PMID: 28397831
  5. Following Schistosoma exposure, TSP-1 levels in the lung increase, via recruitment of circulating monocytes, while TSP-1 inhibition or knockout bone marrow prevents TGF-beta activation and protects against pulmonary hypertension development. PMID: 28555642
  6. TGF-beta/Smad proteins signaling affects radiation response and prolongs survival by regulating DNA repair genes in malignant glioma. PMID: 30230914
  7. The results indicate that EGFR and its activation are critical for YAP-mediated suppression of TGF-beta1-induced apoptosis. This study provides a new understanding of the regulatory mechanism underlying the determination of cell fate in response to TGF-beta1-mediated simultaneous apoptosis and epithelial mesenchymal transformation. PMID: 28425446
  8. transforming growth factor beta (TGFbeta) signaling was upregulated in HSCs from bone marrow of mice with MLL-AF9-induced acute myeloid leukemia (AML) because of excessive production of TGFbeta1, especially from megakaryocytes, and overactivation of latent TGFbeta1 protein. PMID: 29307605
  9. TGF-beta release from platelets is necessary for podoplanin-mediated tumor invasion and metastasis in lung cancer. PMID: 28176852
  10. Data suggest partial or complete transforming growth factor beta 1 (TGFBI) knockdown as a potential therapy against TGFBI-linked corneal dystrophies. PMID: 29117645
  11. De novo formation of the biliary system by TGFbeta-mediated hepatocyte transdifferentiation PMID: 29720662
  12. CXCL9 may promote prostate cancer progression via inhibition of cytokines from T cells. PMID: 29901197
  13. EGCG attenuated airway inflammation in asthmatic mice, decreased the percentage of Th17 cells and increased the percentage of Treg cells. The antiinflammatory effect of EGCG is achieved via the TGFbeta1 signaling pathway. PMID: 29916550
  14. fibrosis induced by Ang II may be alleviated by AKAP12 expression through inactivation of the TGF-beta1 pathway. PMID: 29501491
  15. RUNX1 promoted TGF-beta-induced partial EMT by increasing transcription of the PI3K subunit p110delta, which mediated Akt activation. PMID: 29759484
  16. Transforming growth factor-beta1 induces cerebrovascular dysfunction and astrogliosis through angiotensin II type 1 receptor-mediated signaling pathways PMID: 29505736
  17. TET2 plays an important role in the pathogenesis of diabetic nephropathy by activating TGFbeta1 expression through demethylation of CpG islands in the TGFbeta1 regulatory region PMID: 29705354
  18. early myocardial CTGF mRNA expression (six hours) after Ang-II exposure is likely dependent on latent TGF-beta activation via the canonical Smad-dependent pathway in resident cardiac cells. PMID: 29575960
  19. These results indicate that the hepatocytic expression of TGF-beta and CTGF is mediated by Wnt signalling in Schistosoma japonicum infection. PMID: 28331224
  20. Infection with Mycobacterium bovis results in increase in interleukin-1alpha, TGF-beta1, and MMP1 in multinucleated macrophages. PMID: 29504104
  21. type I and II IFN as in vivo modifiers of LIC fate that may significantly affect the impact of putative leukemia-driving activities such as the ETV6-RUNX1-mediated growth advantage in the presence of TGF-beta and TLR4-upregulated mutagenic activity PMID: 27220664
  22. Studies indicate key developmental processes that require transforming growth factor beta (TGF-beta) signaling [Review]. PMID: 29190317
  23. Osthole,a natural coumarin, decreases the collagen/III ratio in transforming growth factor beta 1 (TGF-beta1)-transfected mouse cardiac fibroblasts via the TGF-beta/Smad signaling pathway. PMID: 29860992
  24. both ERK and Smad2 signal pathways are involved in the activation of macrophages induced by TGF-b1 and high-ambient glucose, while there is no crosstalk shown in the process. PMID: 29199516
  25. In obese mice, periodontitis caused the downregulation of MMP2, and upregulation of TIMP1 and TGF-beta1 at transcriptional and translational levels PMID: 29322806
  26. The protective effect of the EP2 receptor on TGF-beta1 induced podocyte injury via the PI3K / Akt signaling pathway. PMID: 29746568
  27. This study demonstrates that prevention of renal apoB accumulation is a mechanism by which TGF-beta inhibition is nephroprotective. PMID: 28912302
  28. data show that increased TGFbeta in the tumour microenvironment represents a primary mechanism of immune evasion that promotes T-cell exclusion and blocks acquisition of the TH1-effector phenotype; immunotherapies directed against TGFbeta signalling may therefore have broad applications in treating patients with advanced colorectal cancer PMID: 29443964
  29. IL 6 and TGF beta perform essential role in cerebral malaria pathogenesis by modulating the level of glial cell induced neuroinflammation. PMID: 28803696
  30. The increased susceptibility to IMQ-induced psoriasis of GILZ-Tg mice was significantly associated with skin-specific over-activation of TGF-beta1-mediated signaling via SMAD2/3. PMID: 27934944
  31. The data suggest that B cells can down-regulate the function of antigen-presenting cells, and in turn encephalitogenic Th1/Th17 responses, via TGF-beta1. PMID: 27708418
  32. p-SMAD2/3 and p-ERK1/2 might play a regulatory role in TGF-beta1 induced CTGF exp p-SMAD2/3 and p-ERK1/2 might play a regulatory role in TGF-beta1 induced CTGF expression during tooth development. PMID: 28825193
  33. inhibiting NCAM1 would be cardioprotective, counteract the pathological action of TGFbeta1 and reduce heart failure severity. PMID: 28870505
  34. TGF-beta signaling has a role in inhibiting tumor progression and invasion in an induced mouse bladder cancer model PMID: 27378170
  35. calpains inhibition plays crucial roles in vascular restenosis by preventing neointimal hyperplasia at the early stage via suppression of the MMP2/TGF-beta1 pathway. PMID: 27453531
  36. Suggest an essential role for platelet-derived TGFbeta1 for the vascular remodelling response to arterial injury, apparently independent from the role of platelets in thrombosis or haemostasis. PMID: 28726976
  37. In cultured B16 melanoma and Bend3 endothelial cells treated with Bend3 conditioned media, MITF, tyrosinase, and melanin expression decreased due to TGFB1 secreted by the endothelial cells. PMID: 27172887
  38. Lycat regulates TGF-beta mediated lung fibroblast differentiation in pulmonary fibrosis. PMID: 28751023
  39. TFGbeta1 and arachidonic acid synthase expression is increased during the involution period in mammary glands. PMID: 28381667
  40. Chronic exercise suppressed the TGF-beta1/IkB-alpha axis in the hypothalamus and improved the energy homeostasis in an animal model of obesity-associated with aging. PMID: 28854149
  41. miR-146 inhibits skeletal muscle fibrosis by suppressing TGF-beta/Smad4 signaling pathway. PMID: 28510617
  42. Study shows that expression of Gadd45 family members is adjusted in response to KCl/depolarization, BDNF/TRKB, and/or TGFB-signaling. Additionally, results indicate decreased Gadd45 expression in a mouse model of depression, and that Gadd45b expression and DNA demethylation activate Arc transcription. PMID: 28444170
  43. Bioinformatics analysis provides positive supportive correlation between somatic mutations for VD-related genes and the TGF-beta pathway PMID: 27456065
  44. Taken together, these results indicate that eIF6 may be involved in external mechanical force-mediated murine dermal fibroblast function at least partly through the TGF-beta1/TGFBR1/TGFBR2 pathway. PMID: 27824055
  45. important roles in virtually every stage of neural development [review] PMID: 28130363
  46. These data suggest that the interplay between cell-matrix adhesion and intercellular adhesion is an important determinant for some aspects of TGFbeta1-induced epithelial-mesenchymal transition via alphaSMA expression induction. PMID: 27194451
  47. Excessive activation of TGFbeta by spinal instability causes vertebral endplate sclerosis and intervertebral disk degeneration. PMID: 27256073
  48. A Smad3-PTEN regulatory loop controls proliferation and apoptotic responses to TGF-beta in mouse endometrium. PMID: 28524854
  49. TGFss through the Alk1/TgfssR2 receptor axis is acting on endothelial cells to produce hematopoiesis. PMID: 29253505
  50. The adoptive transfer of NK1.1(-) CD4(+) NKG2D(+) cells suppressed DSS-induced colitis largely dependent on TGF-beta. Thus, NK1.1(-) CD4(+) NKG2D(+) cells exhibited immune regulatory functions, and this T cell subset could be developed to suppress inflammation in clinics. PMID: 28224733

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Subcellular Location [Latency-associated peptide]: Secreted, extracellular space, extracellular matrix.; [Transforming growth factor beta-1]: Secreted.
Protein Families TGF-beta family
Database Links

KEGG: mmu:21803

STRING: 10090.ENSMUSP00000002678

UniGene: Mm.248380

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