Revision 2

#77397Store at -20C

1 Kit

(9 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb 19245 20 µl 42 kDa Rabbit IgG
COL1A1 (E8I9Z) Rabbit mAb 91144 20 µl 220 kDa Rabbit IgG
SMAD2/3 (D7G7) XP® Rabbit mAb 8685 20 µl 52, 60 kDa Rabbit IgG
Smad2 (D43B4) XP® Rabbit mAb 5339 20 µl 60 kDa Rabbit IgG
Phospho-SMAD2 (Ser465/Ser467) (E8F3R) Rabbit mAb 18338 20 µl 60 kDa Rabbit IgG
YKL-40 (E2L1M) Rabbit mAb 47066 20 µl 30-40 kDa Rabbit IgG
Phospho-SMAD2 (Ser465/467)/SMAD3 (Ser423/425) (D27F4) Rabbit mAb 8828 20 µl 52, 60 kDa Rabbit IgG
TGF-β (56E4) Rabbit mAb 3709 20 µl 12, 45-60 kDa Rabbit IgG
TGF-β Receptor II (E5M6F) Rabbit mAb 41896 20 µl 85 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

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.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the 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.

  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.
  10. Mack, M. (2018) Matrix Biol 68-69, 106-21.
  11. Johansen, J.S. (2006) Dan Med Bull 53, 172-209.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    CST is a registered trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 5,675,063.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

    Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

    Revision 2
    #77397

    TGF-β Fibrosis Pathway Antibody Sampler Kit

    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 1 Expand Image
    Simple Western™ analysis of extracts (1 mg/mL) from Human Colon tissue using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb #19245. The virtual lane view (left) shows a single target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 2 Expand Image
    Simple WesternTM analysis of lysates (1 mg/ml) from THP-1 cells using YKL-40 (E2L1M) Rabbit mAb #47066. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the JessTM Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230kDa.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 3 Expand Image
    Simple Western™ analysis of lysates (1.0 mg/mL) from serum starved H-1080 cells treated + hTGF-β3 (10ng/mL, 30 mins) using Phospho-SMAD2 (Ser465/467)/SMAD3 (Ser423/425) (D27F4) Rabbit mAb #8828. The virtual lane view (left) shows the target band (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 4 Expand Image
    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).
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 5 Expand Image
    Immunohistochemical analysis of paraffin-embedded human endometrioid carcinoma using alpha-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb performed on the Leica® Bond Rx.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 6 Expand Image
    Western blotting analysis of extracts from various human, mouse, and rat tissues using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb (upper) and GAPDH #5174 (lower). As expected, skeletal muscle samples are negative for α-smooth muscle actin.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 7 Expand Image
    Western blot analysis of recombinant active TGF-β1 using TGF-β (56E4) Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 8 Expand Image
    Western blot analysis of extracts from 293T cells, mock transfected (-), transfected with Myc/DDK-tagged TGF-β Receptor I protein (TGF-β Receptor I-Myc/DDK; +), or transfected with Myc/DDK-tagged TGF-β Receptor II protein (TGF-β Receptor II-Myc/DDK; +), using TGF-β Receptor II (E5M6F) Rabbit mAb (upper), Myc-Tag (71D10) Rabbit mAb #2278 (middle), and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 9 Expand Image
    Western blot analysis of extracts from THP-1 and A-204 using YKL-40 (E2L1M) Rabbit mAb upper) and loading control α-Actinin (D6F6) XP® Rabbit mAb #6487. As expected, A-204 is low to negative for YKL-40 expression.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 10 Expand Image
    Western blot analysis of extracts from HeLa cells (lane 1) or SMAD2 knock-out cells (lane 2) using Smad2 (D43B4) XP® Rabbit mAb #5339 (upper), and α-Actinin (D6F6) XP® Rabbit mAb #6487 (lower). The absence of signal in the SMAD2 knock-out HeLa cells confirms specificity of the antibody for SMAD2.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 11 Expand Image
    After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 12 Expand Image
    Western blot analysis of extracts from HeLa and ACHN cells using SMAD2/3 (D7G7) XP® Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 13 Expand Image
    Western blot analysis of extracts from HaCaT cells, untreated (-) or treated with hTGF-β3 #8425 (+) in the absence or presence of the TGFR inhibitor SB 431542, using Phospho-SMAD2 (Ser465/467)/SMAD3 (Ser423/425) (D27F4) Rabbit mAb (upper) or SMAD2/3 (D7G7) XP® Rabbit mAb #8685 (lower).
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 14 Expand Image
    Western blot analysis of extracts from various cell lines using COL1A1 (E8I9Z) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 15 Expand Image
    Immunoprecipitation of phospho-SMAD2 (Ser465/Ser467) from extracts of HaCaT cells treated with Human Transforming Growth Factor β1 (hTGF-β1) #8915 (10nM, 30mins). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-SMAD2 (Ser465/Ser467) (E8F3R) Rabbit mAb. Western blot analysis was performed using P-SMAD2 (Ser465/ Ser467) (E8F3R) Rabbit mAb. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used for detection to avoid cross-reactivity with IgG.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 16 Expand Image
    Immunoprecipitation of α-smooth muscle actin from mouse colon tissue extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb. Western blot analysis was performed using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 17 Expand Image
    Immunohistochemical analysis of paraffin-embedded human appendix using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 18 Expand Image
    Western blot analysis of extracts of HeLa cells, mock transfected or transfected with TGF-β1 precursor, using TGF-β (56E4) Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 19 Expand Image
    Western blot analysis of extracts from Hep G2 and HuH-7 cells, untreated (-) or treated with PNGase F (+), using TGF-β Receptor II (E5M6F) Rabbit mAb (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 20 Expand Image
    Immunohistochemical analysis of paraffin-embedded human lung squamous cell carcinoma using YKL-40 (E2L1M) Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 21 Expand Image
    Western blot analysis of extracts from various cell lines using Smad2 (D43B4) XP® Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 22 Expand Image
    Immunoprecipitation of SMAD2/3 protein from HCT116 extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is SMAD2/3 (D7G7) XP®. Western blot analysis was performed using SMAD2/3 (D7G7) XP®. Mouse Anti-rabbit IgG (Conformation Specific) (L27A9) mAb #3678 was used as a secondary antibody.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 23 Expand Image
    Immunoprecipitation of COL1A1 protein from U-118 MG cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is COL1A1 (E8I9Z) XP® Rabbit mAb. Western blot analysis was performed using COL1A1 (E8I9Z) XP® Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as secondary antibody.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 24 Expand Image
    Confocal immunofluorescent analysis of serum-starved HT-1080 cells, untreated (left), treated with Human Transforming Growth Factor β3 (hTGF-β3) #8425 (100 ng/mL, 20 min; center), or treated with hTGF-β3 and post-processed with λ-phosphatase (right), using Phospho-Smad2 (Ser465/467) (E8F3R) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red).
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 25 Expand Image
    Immunohistochemical analysis of paraffin-embedded human ductal carcinoma of the breast using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 26 Expand Image
    Western blot analysis of extracts from K-562, Saos-2 and 786-0 cells using TGF-β (56E4) Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 27 Expand Image
    Immunohistochemical analysis of paraffin-embedded human non-Hodgkin's lymphoma using YKL-40 (E2L1M) Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 28 Expand Image
    Confocal immunofluorescent analysis of NIH/3T3 cells, serum-starved (left) or treated with hTGF-β3 #8425 (right), using Smad2 (D43B4) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red).
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 29 Expand Image
    Confocal immunofluorescent analysis of HeLa cells, serum starved (left), treated with hTGF-β3 #8425 (100 ng/ml, 30 min, center), or treated with hTGF-β3 and SB43152 (10 ug/mL, 1 hr, right), using SMAD2/3 (D7G7) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 30 Expand Image
    Flow cytometric analysis of serum-starved HT-1080 cells, untreated (blue), treated with Human Transforming Growth Factor β3 (hTGF-β3) #8425 (100 ng/mL, 30 min; green) or pretreated with SB43152 (10 ug/mL, 30 min) and treated with hTGF-β3 (100 ng/mL, 30 min; red), using Phospho-Smad2 (Ser465/467) (E8F3R) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 31 Expand Image
    Immunohistochemical analysis of paraffin-embedded human colon carcinoma using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 32 Expand Image
    Immunohistochemical analysis of paraffin-embedded human serous papillary carcinoma of the ovary using YKL-40 (E2L1M) Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 33 Expand Image
    Flow cytometric analysis of HeLa cells using Smad2 (D43B4) XP® Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 34 Expand Image
    Flow cytometric analysis of HeLa cells using SMAD2/3 (D7G7) XP® Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary control.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 35 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HaCaT cells treated with Human Growth Factor β1 #8915 (7 ng/ml, 1 hr) and either Phospho-SMAD2 (Ser465/Ser467) (E8F3R) Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ID1 Promoter Primers #5139, Human JunB Promoter Primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 36 Expand Image
    Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 37 Expand Image
    Immunohistochemical analysis of paraffin-embedded THP-1 cell pellet (left, positive) or A-204 cell pellet (right, negative) using YKL-40 (E2L1M) Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 38 Expand Image
    Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 39 Expand Image
    Confocal immunofluorescent analysis of THP-1 cells (left, positive) and A-204 cells (right, negative) using YKL-40 (E2L1M) Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 40 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HaCaT cells treated with Human TGF-β3 #8425 (7 ng/ml) for 1 h and either Smad2 (D43B4) XP® Rabbit mAb #5339 or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human CDKN1A Intron 1 Primers #4669, SimpleChIP® Human ID1 Promoter Primers #5139, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 41 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HaCaT cells treated with hTGF-β3 #8425 (7 ng/ml, 1 hr) and SMAD2/3 (D7G7) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across ID1, a known target gene of SMAD2/3 (see additional figure containing ChIP-qPCR data). For additional ChIP-seq tracks, please download the product datasheet.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 42 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse small intestine using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 43 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HaCaT cells treated with hTGF-β3 #8425 (7 ng/ml, 1 hr) and SMAD2/3 (D7G7) XP® Rabbit mAb, using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. DNA Libraries were prepared using DNA Library Prep Kit for Illumina® (ChIP-seq, CUT&RUN) #56795. The figure shows binding across chromosome 20 (upper), including ID1 (lower), a known target gene of SMAD2/3 (see additional figure containing ChIP-qPCR data).
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 44 Expand Image
    Immunohistochemical analysis of paraffin-embedded mouse skeletal muscle using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 45 Expand Image
    Chromatin immunoprecipitations were performed with cross-linked chromatin from HaCaT cells treated with hTGF-β3 #8425 (7 ng/ml, 1 hr) and either SMAD2/3 (D7G7) XP® Rabbit mAb or Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human CDKN1A Intron 1 Primers #4669, SimpleChIP® Human ID1 Promoter Primers #5139, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 46 Expand Image
    Immunohistochemical analysis of paraffin-embedded rhesus monkey kidney using alpha-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 47 Expand Image
    Immunohistochemical analysis of paraffin-embedded rhesus monkey spleen using alpha-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 48 Expand Image
    Confocal immunofluorescent analysis of mouse small intestine (left) and skeletal muscle (right) using α-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb (green). Actin filaments were labeled with DyLight 554 Phalloidin #13054 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 49 Expand Image
    Immunohistochemical analysis of paraffin-embedded rat colon using alpha-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 50 Expand Image

    Immunohistochemical analysis of paraffin-embedded rat brain using alpha-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.

    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 51 Expand Image
    Immunohistochemical analysis of paraffin-embedded Syrian hamster ovary using alpha-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.
    TGF-β Fibrosis Pathway Antibody Sampler Kit: Image 52 Expand Image
    Immunohistochemical analysis of paraffin-embedded Syrian hamster uterus using alpha-Smooth Muscle Actin (D4K9N) XP® Rabbit mAb.