使用敲除细胞株进行验证重组RabMAb

重组Anti-Smad3抗体[EP568Y] (ab40854)

Reviews (9)Q&A (1)References (150)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP568Y] to Smad3
  • Suitable for: WB, IHC-P, Sandwich ELISA, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

概述

  • 产品名称

    Anti-Smad3抗体[EP568Y]
    参阅全部 Smad3 一抗

  • 描述

    兔单克隆抗体[EP568Y] to Smad3

  • 宿主

    Rabbit

  • 特异性

    This antibody is specific for Smad3.

  • 经测试应用

    适用于: WB, IHC-P, Sandwich ELISA, Flow Cyt, ICC/IFmore details
    不适用于: IP

  • 种属反应性

    与反应: Mouse, Rat, Human
    预测可用于: Chicken, Pig

  • 免疫原

    Synthetic peptide within Human Smad3 aa 200-300 (internal sequence). The exact sequence is proprietary.
    Database link: P84022
    (Peptide available as ab173094)

  • 阳性对照

    • WB: A549, HeLa, Human Kidney, HT-29, HT-1080 and Jurkat whole cell lysates; Rat liver tissue lysate. IHC-P: Human prostate carcinoma, breast carcinoma, colonic adenocarcinoma, lung adenocarcinoma, gastric adenocarcinoma, glioma and liver tissues. ICC/IF: HepG2 cells. Flow Cyt: HCT116 and HT-29 cells.

  • 常规说明

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

性能

应用

Our Abpromise guarantee covers the use of ab40854 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/1000 - 1/10000. Detects a band of approximately 55 kDa (predicted molecular weight: 48 kDa).

 
IHC-P 1/500 - 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.
Sandwich ELISA Use a concentration of 0.5 µg/ml.

For sandwich ELISA, use this antibody as Detection at 0.5 µg/ml with Mouse monoclonal [AF9F7] to Smad3 (ab75512) as Capture.
Flow Cyt 1/50 - 1/210.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF 1/500 - 1/2000.

 
  • 应用说明
    Is unsuitable for IP.

    • 靶标

    • 功能

      Receptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes that are regulated by TGF-beta and, on formation of the SMAD3/SMAD4 complex, activates transcription. Also can form a SMAD3/SMAD4/JUN/FOS complex at the AP-1/SMAD site to regulate TGF-beta-mediated transcription. Has an inhibitory effect on wound healing probably by modulating both growth and migration of primary keratinocytes and by altering the TGF-mediated chemotaxis of monocytes. This effect on wound healing appears to be hormone-sensitive. Regulator of chondrogenesis and osteogenesis and inhibits early healing of bone fractures. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator.

    • 疾病相关

      Colorectal cancer
      Loeys-Dietz syndrome 3

    • 序列相似性

      Belongs to the dwarfin/SMAD family.
      Contains 1 MH1 (MAD homology 1) domain.
      Contains 1 MH2 (MAD homology 2) domain.

    • 结构域

      The MH1 domain is required for DNA binding. Also binds zinc ions which are necessary for the DNA binding.
      The MH2 domain is required for both homomeric and heteromeric interactions and for transcriptional regulation. Sufficient for nuclear import.
      The linker region is required for the TGFbeta-mediated transcriptional activity and acts synergistically with the MH2 domain.

    • 翻译后修饰

      Phosphorylated on serine and threonine residues. Enhanced phosphorylation in the linker region on Thr-179, Ser-204 and Ser-208 on EGF and TGF-beta treatment. Ser-208 is the main site of MAPK-mediated phosphorylation. CDK-mediated phosphorylation occurs in a cell-cycle dependent manner and inhibits both the transcriptional activity and antiproliferative functions of SMAD3. This phosphorylation is inhibited by flavopiridol. Maximum phosphorylation at the G(1)/S junction. Also phosphorylated on serine residues in the C-terminal SXS motif by TGFBR1 and ACVR1. TGFBR1-mediated phosphorylation at these C-terminal sites is required for interaction with SMAD4, nuclear location and transactivational activity, and appears to be a prerequisite for the TGF-beta mediated phosphorylation in the linker region. Dephosphorylated in the C-terminal SXS motif by PPM1A. This dephosphorylation disrupts the interaction with SMAD4, promotes nuclear export and terminates TGF-beta-mediated signaling. Phosphorylation at Ser-418 by CSNK1G2/CK1 promotes ligand-dependent ubiquitination and subsequent proteasome degradation, thus inhibiting SMAD3-mediated TGF-beta responses. Phosphorylated by PDPK1.
      Acetylation in the nucleus by EP300 in the MH2 domain regulates positively its transcriptional activity and is enhanced by TGF-beta.
      Ubiquitinated. Monoubiquitinated, leading to prevent DNA-binding. Deubiquitination by USP15 alleviates inhibition and promotes activation of TGF-beta target genes.
      Poly-ADP-ribosylated by PARP1 and PARP2. ADP-ribosylation negatively regulates SMAD3 transcriptional responses during the course of TGF-beta signaling.

    • 细胞定位

      Cytoplasm. Nucleus. Cytoplasmic and nuclear in the absence of TGF-beta. On TGF-beta stimulation, migrates to the nucleus when complexed with SMAD4 (PubMed:15799969). Through the action of the phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out of the nucleus by interaction with RANBP1 (PubMed:16751101, PubMed:19289081). Co-localizes with LEMD3 at the nucleus inner membrane (PubMed:15601644). MAPK-mediated phosphorylation appears to have no effect on nuclear import (PubMed:19218245). PDPK1 prevents its nuclear translocation in response to TGF-beta (PubMed:17327236).
    • Information by UniProt

    • 数据库链接

      see all

    • 别名

      • DKFZP586N0721 antibody
      • DKFZp686J10186 antibody
      • hMAD 3 antibody
      • hMAD-3 antibody
      • hSMAD3 antibody
      • HSPC193 antibody
      • HST17436 antibody
      • JV15 2 antibody
      • JV15-2 antibody
      • JV152 antibody
      • LDS1C antibody
      • LDS3 antibody
      • MAD (mothers against decapentaplegic Drosophila) homolog 3 antibody
      • MAD homolog 3 antibody
      • Mad homolog JV15 2 antibody
      • Mad protein homolog antibody
      • MAD, mothers against decapentaplegic homolog 3 antibody
      • Mad3 antibody
      • MADH 3 antibody
      • MADH3 antibody
      • MGC60396 antibody
      • Mothers against decapentaplegic homolog 3 antibody
      • Mothers against DPP homolog 3 antibody
      • SMA and MAD related protein 3 antibody
      • SMAD 3 antibody
      • SMAD antibody
      • SMAD family member 3 antibody
      • SMAD, mothers against DPP homolog 3 antibody
      • Smad3 antibody
      • SMAD3_HUMAN antibody
      see all

    图片

    • Western blot - Anti-Smad3 antibody [EP568Y] (ab40854)
      Western blot - Anti-Smad3 antibody [EP568Y] (ab40854)
      All lanes : Anti-Smad3 antibody [EP568Y] (ab40854) at 1/1000 dilution

      Lane 1 : Wild-type A549 cell lysate
      Lane 2 : SMAD3 knockout A549 cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : Human Kidney cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 48 kDa
      Observed band size: 50 kDa
      why is the actual band size different from the predicted?



      Lanes 1 - 4: Merged signal (red and green). Green - ab40854 observed at 48 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

      ab40854 was shown to react with Smad3 in western blot. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab40854 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad3 antibody [EP568Y] (ab40854)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad3 antibody [EP568Y] (ab40854)

      Immunohistochemical analysis of paraffin-embedded human colonic adenocarcinoma tissue labelling Smad3 with unpurified ab40854.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-Smad3 antibody [EP568Y] (ab40854)
      Immunocytochemistry/ Immunofluorescence - Anti-Smad3 antibody [EP568Y] (ab40854)

      Immunocytochemsitry/Immunofluorescence analysis of HepG2 cells labelling Smad3 (green) with purified ab40854 at 1/2000 . Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody (green, left panel). Counterstained with DAPI (blue, right panel).

    • Flow Cytometry - Anti-Smad3 antibody [EP568Y] (ab40854)
      Flow Cytometry - Anti-Smad3 antibody [EP568Y] (ab40854)

      Overlay histogram showing HCT116 cells stained with unpurified ab40854 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab40854, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    • Sandwich ELISA - Anti-Smad3 antibody [EP568Y] (ab40854)
      Sandwich ELISA - Anti-Smad3 antibody [EP568Y] (ab40854)

      Standard Curve for Smad3 (Analyte: Smad3 protein (His tag) (ab89353, unpurified)); dilution range 1pg/ml to 1µg/ml using Capture Antibody Mouse monoclonal [AF9F7] to Smad3 (ab75512) at 5µg/ml and Detector Antibody Rabbit monoclonal [EP568Y] to Smad3 (ab40854) at 0.5µg/ml.

    • Immunocytochemistry/ Immunofluorescence - Anti-Smad3 antibody [EP568Y] (ab40854)
      Immunocytochemistry/ Immunofluorescence - Anti-Smad3 antibody [EP568Y] (ab40854)This image is courtesy of an Abreview submitted by Francesco Elia Marino

      ab40854 staining Smad3 in human granulosa cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with ethanol and triton and blocked for 1 hour at 26°C. Samples were incubated with primary antibody (1/200) for 16 hours at 4°C. An undiluted IRDye® 800CW-conjugated goat anti-rabbit IgG (H+L) polyclonal was used as the secondary antibody. Left - negative control (4 replicates).

      See Abreview

    • Western blot - Anti-Smad3 antibody [EP568Y] (ab40854)
      Western blot - Anti-Smad3 antibody [EP568Y] (ab40854)
      All lanes : Anti-Smad3 antibody [EP568Y] (ab40854) at 1/5300 dilution (purified)

      Lane 1 : HT-29 cell lysate
      Lane 2 : HT-1080 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 48 kDa
      Observed band size: 58 kDa why is the actual band size different from the predicted?



      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Western blot - Anti-Smad3 antibody [EP568Y] (ab40854)
      Western blot - Anti-Smad3 antibody [EP568Y] (ab40854)
      Anti-Smad3 antibody [EP568Y] (ab40854) at 1/5000 dilution (unpurified) + Jurkat cell lysate at 10 µg

      Predicted band size: 48 kDa
      Observed band size: 55 kDa why is the actual band size different from the predicted?

    • Western blot - Anti-Smad3 antibody [EP568Y] (ab40854)
      Western blot - Anti-Smad3 antibody [EP568Y] (ab40854)
      Anti-Smad3 antibody [EP568Y] (ab40854) at 1/5300 dilution (purified) + Rat liver tissue lysate at 10 µg

      Secondary
      Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

      Predicted band size: 48 kDa
      Observed band size: 58 kDa why is the actual band size different from the predicted?



      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • Flow Cytometry - Anti-Smad3 antibody [EP568Y] (ab40854)
      Flow Cytometry - Anti-Smad3 antibody [EP568Y] (ab40854)

      Flow cytometry analysis of HT-29 cells labelling Smad3 with purified ab40854 at 1/210 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad3 antibody [EP568Y] (ab40854)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad3 antibody [EP568Y] (ab40854)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling Smad3 with purified ab40854 at 1/2000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad3 antibody [EP568Y] (ab40854)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad3 antibody [EP568Y] (ab40854)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling unpurified ab40854 at 1/100 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad3 antibody [EP568Y] (ab40854)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad3 antibody [EP568Y] (ab40854)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling unpurified ab40854.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad3 antibody [EP568Y] (ab40854)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad3 antibody [EP568Y] (ab40854)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung adenocarcinoma tissue labelling unpurified ab40854.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad3 antibody [EP568Y] (ab40854)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad3 antibody [EP568Y] (ab40854)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human glioma tissue labelling unpurified ab40854.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad3 antibody [EP568Y] (ab40854)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Smad3 antibody [EP568Y] (ab40854)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric adenocarcinoma tissue labelling unpurified ab40854.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Anti-Smad3 antibody [EP568Y] (ab40854)
      Anti-Smad3 antibody [EP568Y] (ab40854)

    文献 (150)

    发表研究结果有使用 ab40854？请让我们知道，以便我们可以引用本数据表中的参考文章。

    ab40854 被引用在 150 文献中.

    • Qu S  et al. MicroRNA-194 reduces inflammatory response and human dermal microvascular endothelial cells permeability through suppression of TGF-ß/SMAD pathway by inhibiting THBS1 in chronic idiopathic urticaria. J Cell Biochem 121:111-124 (2020). PubMed: 31190349
    • Guo S  et al. Inhibition mechanism of lung cancer cell metastasis through targeted regulation of Smad3 by miR-15a. Oncol Lett 19:1516-1522 (2020). PubMed: 31966076
    • Chen X  et al. Exosome-mediated transfer of miR-93-5p from cancer-associated fibroblasts confer radioresistance in colorectal cancer cells by downregulating FOXA1 and upregulating TGFB3. J Exp Clin Cancer Res 39:65 (2020). PubMed: 32293494
    • Zhang Z  et al. Aspirin inhibits endometrial fibrosis by suppressing the TGF-ß1-Smad2/Smad3 pathway in intrauterine adhesions. Int J Mol Med 45:1351-1360 (2020). PubMed: 32323728
    • Xiao Z  et al. RUNX3 inhibits the invasion and migration of esophageal squamous cell carcinoma by reversing the epithelial-mesenchymal transition through TGF-ß/Smad signaling. Oncol Rep 43:1289-1299 (2020). PubMed: 32323849
    View all Publications for this product

    客户评价及客户问答

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    1-10 of 10 Abreviews or Q&A

    Western blot abreview for Anti-Smad3 antibody [EP568Y]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (melanoma)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    20 µg
    Treatment
    TFG-beta1 for 1hr
    Specification
    melanoma
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    提交于 Jul 23 2019

    Western blot abreview for Anti-Smad3 antibody [EP568Y]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - whole (Muscle)
    Gel Running Conditions
    Reduced Denaturing (9)
    Loading amount
    50 µg
    Specification
    Muscle
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Mr. Baptiste Jude

    Verified customer

    提交于 Feb 28 2018

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Smad3 antibody [EP568Y]

     Excellent
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Lung tissue)
    Antigen retrieval step
    None
    Permeabilization
    No
    Specification
    Lung tissue
    Blocking step
    Zytomed Systems HRP Polymer Blocking Reagent as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 25°C
    Fixative
    HOPE-fixation
    Read More

    Abcam user community

    Verified customer

    提交于 Oct 06 2015

    Western blot abreview for Anti-Smad3 antibody [EP568Y]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Saos)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    20 µg
    Specification
    Saos
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5µg/mL · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    提交于 Apr 21 2015

    Western blot abreview for Anti-Smad3 antibody [EP568Y]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    20 µg
    Gel Running Conditions
    Reduced Denaturing
    Sample
    Human Cell lysate - whole cell (Normal Human Keratinocyte)
    Specification
    Normal Human Keratinocyte
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Abcam user community

    Verified customer

    提交于 Mar 30 2015

    Western blot abreview for Anti-Smad3 antibody [EP568Y]

     Good
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    50 µg
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Sample
    Human Cell lysate - whole cell (HeLa)
    Specification
    HeLa
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More

    Abcam user community

    Verified customer

    提交于 Oct 24 2014

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Smad3 antibody [EP568Y]

     Excellent
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C
    Sample
    Human Cell (Human granulosa cell Line)
    Specification
    Human granulosa cell Line
    Permeabilization
    Yes - Ethanol and triton
    Fixative
    Paraformaldehyde
    Read More

    Dr. Francesco Elia Marino

    Verified customer

    提交于 Jun 09 2014

    Western blot abreview for Anti-Smad3 antibody [EP568Y]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    40 µg
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (12)
    Sample
    Mouse Tissue lysate - whole (Gastrocnemius (Muscle))
    Specification
    Gastrocnemius (Muscle)
    Blocking step
    Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 27°C
    Read More

    Dr. Francesco Elia Marino

    Verified customer

    提交于 Mar 07 2014

    Question

    Hello, I'm looking for antibodies that would let me distinguish Smad2 and Smad3 in paraffin sections of mouse heart. What would you recommend?

    Read More
    Answer

    Thank you for contacting us.

    We do have a number of anti-SMAD2 and anti-SMAD3 antibodies which we guarantee in IHC-P on mouse. We to don't have the opportunity to test specific products against one another in each application but based on Abreviews, publications and other data available on our datasheets I can recommend the following products.

    For identifying both SMAD2 and SMAD3 in IHC-P https://www.abcam.com/Smad2-Smad3-antibody-ab65847.html https://www.abcam.com/Smad2-Smad3-antibody-ab65847.html is guaranteed by our Abpromise in mouse.

    While we have not tested our SMAD2 products for SMAD3 reactivity or the SMAD3 against SMAD2, I can recommend the following products for identifying the specific proteins based on sequence homologies.

    For identifying only SMAD2 in IHC-P we recommend the rabbit monoclonal https://www.abcam.com/Smad2-antibody-EP567Y-ab33875.html https://www.abcam.com/Smad2-antibody-EP567Y-ab33875.html. The immunogen used for this product has only a 44.4% identity to SMAD3 Mouse.

    For identifying only SMAD2 in IHC-P we recommend the rabbit monoclonal https://www.abcam.com/Smad3-antibody-EP568Y-ab40854.html https://www.abcam.com/Smad3-antibody-EP568Y-ab40854.html. The immunogen used for this product has only a 47.4% identity to SMAD2 Mouse.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Use our products? Submit an Abreview. Earn rewards!
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    Western blot abreview for Anti-Smad3 antibody [EP568Y]

     Good
    Abreviews
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Hela Cells)
    Loading amount
    30 µg
    Specification
    Hela Cells
    Gel Running Conditions
    Non-reduced Denaturing (10% gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More

    Abcam user community

    Verified customer

    提交于 Jun 13 2011

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