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TGF-beta Fibrosis Pathway Antibody Kit

RMB: 5300

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Catalog# HAK21004

TGF-beta Fibrosis Pathway Antibody Kit

Application

  • WB

Reactivity

  • Human

General Overview

Kit Components

Product Includes Specification Application Reactivity Mw
alpha smooth muscle Actin[ET1607-53] 20µl WB,IF-Cell,IF-Tissue,IHC-P,FC,mIHC Human,Mouse,Rat Predicted band size: 42 kDa
COL1A1[HA722517] 20µl WB,IHC-P,IF-Tissue,IF-Cell,FC,IP,mIHC,IHC-Fr Human,Mouse,Rat Predicted band size: 139 kDa
Smad2+Smad3[HA724174] 20µl
Smad2[ET1604-22] 20µl WB,IF-Cell,IHC-P,IP,FC Human,Mouse,Rat Predicted band size: 52 kDa
Phospho-SMAD2 (S465+467)[HA722443] 20µl WB,IF-Cell,IHC-P Human,Mouse,Rat Predicted band size: 52/48 kDa
YKL-40 / CHI3L1[EM1902-14] 20µl WB,IHC-P,IF-Cell,FC,IF-Tissue Human,Mouse Predicted band size: 43 kDa
TGF beta Receptor II[HA721693] 20µl WB Human Predicted band size: 65 kDa
TGF beta 1[HA721143] 20µl WB,IHC-P,IF-Tissue Human,Mouse,Rat,Monkey Predicted band size: 44 kDa
Goat Anti-Rabbit IgG (H+L)[HA1001] 100µl WB,ELISA,IHC-P Rabbit
Goat Anti-Mouse IgG (H+L)[HA1006] 100µl WB,ELISA,IHC-P Mouse

Product Description

Transforming growth factor- β The fibrosis pathway Antibody Sampler Kit provides an economical method to study the activation of transforming growth factor- β/ Smad2 / 3 signaling pathway leads to the expression of Pro fibrogenic genes in cells or tissues, including alpha smooth muscle Actin, COL1A1/Collagen Ⅰ and YKL-40 / CHI3L1were up-regulated in activated fibroblasts. The kit contains enough antibodies to perform at least two Western blotting tests on each primary antibody.And also includes secondary reagent for detection of these antibodies.

Product Features

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Background

Transforming growth factor-β (TGF-β) superfamily members are critical regulators of cell proliferation and differentiation, developmental patterning and morphogenesis, and disease pathogenesis.</br> In the context of fibrosis, TGF-β signaling to SMAD2/3 is one of the biggest drivers of the profibrotic program.TGF-β elicits signaling through three cell surface receptors: type I (RI), type II (RII), and type III (RIII).Activated type I receptors associate with SMAD2/3 and phosphorylate them on a conserved carboxy terminal SSXS motif.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. TGF-β/ SMAD2/3 signaling also induces expression of α-Smooth Muscle Actin in fibroblasts, causing transformation of these cells to myofibroblasts. Injury to the tissue attracts macrophages and other immune cells and the fibrotic tissue soon becomes a site of inflammation. 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.

Data Links

Background References

1. Massagué, J. et al. (2000) Cell 103, 295-309.

2. de Caestecker, M.P. et al. (2000) J Natl Cancer Inst 92, 1388-402.

3. Derynck, R. et al. (2001) Nat Genet 29, 117-29.

4. Miyazono, K. et al. (2000) Adv Immunol 75, 115-57.

5. Meng, X.M. et al. (2016) Nat Rev Nephrol 12, 325-38.

6. Wu, G. et al. (2000) Science 287, 92-7.

7. Attisano, L. and Wrana, J.L. (2002) Science 296, 1646-7.

8. Moustakas, A. et al. (2001) J Cell Sci 114, 4359-69.

9. Bagalad, B.S. et al. J Oral Maxillofac Pathol 21, 462-3.

Images

  • Western blot analysis of alpha smooth muscle Actin on different lysates with Rabbit anti-alpha smooth muscle Actin antibody (<a href="/products/ET1607-53" style="font-weight: bold;text-decoration: underline;">ET1607-53</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: A431 cell lysate<br />Lane 3: A549 cell lysate<br />Lane 4: NIH/3T3 cell lysate<br />Lane 5: C2C12 cell lysate<br />Lane 6: L6 cell lysate<br />Lane 7: Mouse heart tissue lysate<br />Lane 8: Mouse skin tissue lysate<br />Lane 9: Rat heart tissue lysate<br />Lane 10: Rat skin tissue lysate<br />Lane 11: Rat smooth muscle tissue lysate<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 42 kDa<br />Observed band size: 42 kDa<br /><br />Exposure time: 5 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1607-53" style="font-weight: bold;text-decoration: underline;">ET1607-53</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of alpha smooth muscle Actin on different lysates with Rabbit anti-alpha smooth muscle Actin antibody (ET1607-53) at 1/5,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: A431 cell lysate
    Lane 3: A549 cell lysate
    Lane 4: NIH/3T3 cell lysate
    Lane 5: C2C12 cell lysate
    Lane 6: L6 cell lysate
    Lane 7: Mouse heart tissue lysate
    Lane 8: Mouse skin tissue lysate
    Lane 9: Rat heart tissue lysate
    Lane 10: Rat skin tissue lysate
    Lane 11: Rat smooth muscle tissue lysate

    Lysates/proteins at 20 µg/Lane.

    Predicted band size: 42 kDa
    Observed band size: 42 kDa

    Exposure time: 5 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1607-53) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Smad2 on different lysates with Rabbit anti-Smad2 antibody (<a href="/products/ET1604-22" style="font-weight: bold;text-decoration: underline;">ET1604-22</a>) at 1/5,000 dilution.<br /><br />Lane 1: HeLa cell lysate<br />Lane 2: HT-29 cell lysate<br />Lane 3: Jurkat cell lysate<br />Lane 4: HL-60 cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 52 kDa<br />Observed band size: 58 kDa<br /><br />Exposure time: 1 minute 20 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1604-22" style="font-weight: bold;text-decoration: underline;">ET1604-22</a>) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Smad2 on different lysates with Rabbit anti-Smad2 antibody (ET1604-22) at 1/5,000 dilution.

    Lane 1: HeLa cell lysate
    Lane 2: HT-29 cell lysate
    Lane 3: Jurkat cell lysate
    Lane 4: HL-60 cell lysate

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 52 kDa
    Observed band size: 58 kDa

    Exposure time: 1 minute 20 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1604-22) at 1/5,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of Phospho-Smad3(S423/S425) on different lysates with Rabbit anti-Phospho-Smad3(S423/S425) antibody (<a href="/products/ET1609-41" style="font-weight: bold;text-decoration: underline;">ET1609-41</a>) at 1/2,000 dilution.<br /><br />Lane 1: A549 whole cell lysate<br />Lane 2: A549 treated with 5ng/mL TGF-beta1 for 24 hours whole cell lysate<br />Lane 3: C2C12 whole cell lysate<br />Lane 4: C2C12 treated with 5ng/mL TGF-beta1 for 24 hours whole cell lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br /><br />Predicted band size: 48 kDa<br />Observed band size: 55 kDa<br /><br />Exposure time: 3 minutes 30 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1609-41" style="font-weight: bold;text-decoration: underline;">ET1609-41</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1:50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Phospho-Smad3(S423/S425) on different lysates with Rabbit anti-Phospho-Smad3(S423/S425) antibody (ET1609-41) at 1/2,000 dilution.

    Lane 1: A549 whole cell lysate
    Lane 2: A549 treated with 5ng/mL TGF-beta1 for 24 hours whole cell lysate
    Lane 3: C2C12 whole cell lysate
    Lane 4: C2C12 treated with 5ng/mL TGF-beta1 for 24 hours whole cell lysate

    Lysates/proteins at 15 µg/Lane.

    Predicted band size: 48 kDa
    Observed band size: 55 kDa

    Exposure time: 3 minutes 30 seconds;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1609-41) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

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