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TGF-β Fibrosis Pathway Antibody Sampler Kit #77397

Western blot analysis of extracts from HaCaT cells, untreated (-) or treated with Human Transforming Growth Factor β3 (hTGF-β3) #8425 (100ng/ml, 30mins) (+), using Phospho SMAD2 (Ser465/Ser467) (E8F3R) Rabbit mAb (upper) and total Smad2 (D43B4) XP® Rabbit mAb, #5339 (lower).

To Purchase # 77397

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Product IncludesQuantityApplicationsReactivityMW(kDa)Isotype
α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb #1924520 µlWB, IP, IHC, IFH M R Hm Mk42Rabbit IgG
COL1A1 (E8I9Z) Rabbit mAb #9114420 µlWB, IPH220Rabbit IgG
SMAD2/3 (D7G7) XP® Rabbit mAb #868520 µlWB, IP, IF, F, ChIPH M R Mk52, 60Rabbit IgG
SMAD2 (D43B4) XP® Rabbit mAb #533920 µlWB, IP, IF, F, ChIPH M R Mk60Rabbit IgG
Phospho-SMAD2 (Ser465/Ser467) (E8F3R) Rabbit mAb #1833820 µlWB, IP, IF, F, ChIPH M R60Rabbit IgG
YKL-40 (E2L1M) Rabbit mAb #4706620 µlWB, IHC, IFH30-40Rabbit IgG
Phospho-SMAD2 (Ser465/467)/SMAD3 (Ser423/425) (D27F4) Rabbit mAb #882820 µlWBH M R Mk52, 60Rabbit IgG
TGF-β (56E4) Rabbit mAb #370920 µlWBH12, 45-60Rabbit IgG
TGF-β Receptor II (E5M6F) Rabbit mAb #4189620 µlWBH85Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074100 µlWBRabGoat 

Product Information

Product Description

The TGF-β Fibrosis Pathway Antibody Sampler Kit provides an economical means of investigating activation of TGF-β/ SMAD2/3 signaling pathways in cells or tissues that lead to the expression of profibrotic genes, including expression of α-Smooth Muscle Actin in activated fibroblasts, and upregulation of Collagen1A1, Col11A1, and YKL-40. The kit includes enough antibodies to perform at least two western blot experiments with each primary antibody.

Background

Transforming growth factor-β (TGF-β) superfamily members are critical regulators of cell proliferation and differentiation, developmental patterning and morphogenesis, and disease pathogenesis (1-4). In the context of fibrosis, TGF-β signaling to SMAD2/3 is one of the biggest drivers of the profibrotic program (5).

TGF-β elicits signaling through three cell surface receptors: type I (RI), type II (RII), and type III (RIII). In response to ligand binding, the type II receptors form stable heterotrimeric complexes with the type I receptors, allowing phosphorylation and activation of type I receptor kinase. Activated type I receptors associate with SMAD2/3 and phosphorylate them on a conserved carboxy terminal SSXS motif. The phosphorylated SMADs dissociate from the receptor and form a heterotrimeric complex with the co-Smad (Smad4), allowing translocation of the complex to the nucleus. Once in the nucleus, phosphorylated SMAD2/3 targets a subset of DNA binding proteins to regulate the transcriptional program (6-8).

In the context of fibrosis, SMAD2/3 activation upregulates expression of profibrotic genes such as COL1A1 and other ECM modulators that modify the extracellular matrix of the tissue. (9). TGF-β/ SMAD2/3 signaling also induces expression of α-Smooth Muscle Actin in fibroblasts, causing transformation of these cells to myofibroblasts (10). Myofibroblasts further modify the ECM, causing excessive accumulation of collagens and other ECM components. Injury to the tissue attracts macrophages and other immune cells and the fibrotic tissue soon becomes a site of inflammation (11). In this pro-fibrotic, pro-inflammatory environment, YKL-40, also known as Chitinase-3-like protein 1 (CHI3L1), is secreted. YKL-40 is a pro-inflammatory glycoprotein that also contributes to the progression of fibrosis (12). Measurement of collagen content, α-Smooth Muscle Actin, and the release of YKL-40 are predictive of fibrotic activity.

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For Research Use Only. Not for Use in Diagnostic Procedures.
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U.S. Patent No. 5,675,063.
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