概述

  • 产品名称Anti-Smad2抗体[EP784Y]
    参阅全部 Smad2 一抗
  • 描述
    兔单克隆抗体[EP784Y] to Smad2
  • 特异性This antibody is specific for MH 1 domain of Smad2.
  • 经测试应用适用于: IHC-P, IP, ICC/IF, WB, Flow Cytmore details
  • 种属反应性
    与反应: Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive)

  • 阳性对照
    • Jurkat cell lysate and human prostate carcinoma tissue.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

    The rat recommendation is based on the WB results.This antibody may not be suitable for IHC with rat samples.

    Mouse: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Produced using Abcam's RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5, 675, 063 and/or 7, 429, 487.

    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.

性能

应用

Our Abpromise guarantee covers the use of ab40855 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
IHC-P 1/50.
IP 1/20 - 1/50.
ICC/IF 1/100 - 1/250.
WB 1/2000 - 1/10000. Detects a band of approximately 55 kDa (predicted molecular weight: 58 kDa).
Flow Cyt 1/20 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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 SMAD2/SMAD4 complex, activates transcription. May act as a tumor suppressor in colorectal carcinoma.
  • 组织特异性Expressed at high levels in skeletal muscle, heart and placenta.
  • 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.
  • 翻译后修饰Phosphorylated on one or several of Thr-220, Ser-245, Ser-250, and Ser-255. In response to TGF-beta, phosphorylated on Ser-465/467 by TGF-beta and activin type 1 receptor kinases. Able to interact with SMURF2 when phosphorylated on Ser-465/467, recruiting other proteins, such as SNON, for degradation. In response to decorin, the naturally occurring inhibitor of TGF-beta signaling, phosphorylated on Ser-240 by CaMK2. Phosphorylated by MAPK3 upon EGF stimulation; which increases transcriptional activity and stability, and is blocked by calmodulin.
    In response to TGF-beta, ubiquitinated by NEDD4L; which promotes its degradation.
    Acetylated on Lys-19 by coactivators in response to TGF-beta signaling, which increases transcriptional activity. Isoform short: Acetylation increases DNA binding activity in vitro and enhances its association with target promoters in vivo. Acetylation in the nucleus by EP300 is enhanced by TGF-beta.
  • 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. On dephosphorylation by phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out of the nucleus by interaction with RANBP1.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Drosophila, homolog of, MADR2 antibody
    • hMAD-2 antibody
    • HsMAD2 antibody
    • JV18 antibody
    • JV18-1 antibody
    • JV181 antibody
    • MAD antibody
    • MAD homolog 2 antibody
    • MAD Related Protein 2 antibody
    • Mad-related protein 2 antibody
    • MADH2 antibody
    • MADR2 antibody
    • MGC22139 antibody
    • MGC34440 antibody
    • Mother against DPP homolog 2 antibody
    • Mothers against decapentaplegic homolog 2 antibody
    • Mothers against decapentaplegic, Drosophila, homolog of, 2 antibody
    • Mothers against DPP homolog 2 antibody
    • OTTHUMP00000163489 antibody
    • Sma and Mad related protein 2 antibody
    • Sma- and Mad-related protein 2 MAD antibody
    • SMAD 2 antibody
    • SMAD family member 2 antibody
    • SMAD, mothers against DPP homolog 2 antibody
    • SMAD2 antibody
    • SMAD2_HUMAN antibody
    see all

Anti-Smad2 antibody [EP784Y] 图像

  • All lanes : Anti-Smad2 antibody [EP784Y] (ab40855) at 1/2000 dilution

    Lane 1 : A-673 (Human muscle Ewing's Sarcoma cell line) whole cell lysate
    Lane 2 : HUVEC (Human umbilical vein endothelial cell line) whole cell lysate
    Lane 3 : C6 (Rat glial tumor cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size : 58 kDa

    Diluting and blocking buffer: 5% NFDM /TBST

  • ab40855 staining Smad2 in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/100. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. ab195889 was used as a counterstain for primary antibody ab40855 at 1/1000. DAPI was used as a nuclear counterstain and PBS as a negative control.

  • All lanes : Anti-Smad2 antibody [EP784Y] (ab40855) at 1/10000 dilution

    Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 20000 µg (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size : 58 kDa

    Blocking and diluting buffer: 5% NFDM/TBST

  • ab40855 stainingSmad2 in human bladder carcinoma tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/50. A ImmunoHistoProbe one step HRP Polymer was used as a secondary antibody, ready to use.

    Negative control 1: PBS in place of primary antibody.

  • ab40855 staining Smad2 in the human cell line HeLa (Human epithelial cell line from cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • Immunofluorescence staining of HeLa cells with purified ab40855 at a working dilution of 1/500, counter-stained with DAPI. The secondary antibody was an Alexa Fluor® 488 conjugated goat anti-rabbit (ab150077), used at a dilution of 1/1000. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, PBS was used instead of the primary antibody.

  • Overlay histogram showing PC3 cells stained with ab40855 (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 (ab40855, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • Anti-Smad2 antibody [EP784Y] (ab40855) at 1/500000 dilution + Jurkat cell lysate

    Predicted band size : 58 kDa
    Observed band size : 58 kDa
  • ab40855 at a 1:100 dilution staining Smad2 in human prostate carcinoma tissue.

Anti-Smad2 antibody [EP784Y] (ab40855)参考文献

This product has been referenced in:
  • Ungefroren H  et al. Rac1b negatively regulates TGF-ß1-induced cell motility in pancreatic ductal epithelial cells by suppressing Smad signalling. Oncotarget 5:277-90 (2014). WB ; Human . Read more (PubMed: 24378395) »
  • Harazono Y  et al. miR-655 Is an EMT-suppressive MicroRNA targeting ZEB1 and TGFBR2. PLoS One 8:e62757 (2013). WB ; Human . Read more (PubMed: 23690952) »

See all 7 Publications for this product

Product Wall

Application Western blot
Loading amount 60 µg
Gel Running Conditions Reduced Denaturing (4-20% mini gel)
Sample Mouse Tissue lysate - whole (Penile Tissue)
Specification Penile Tissue
Blocking step SuperBlock Blocking Buffer (PBS) as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Miss. gwen Lagoda

Verified customer

提交于 Nov 12 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (Penile Tissue)
Loading amount 60 µg
Specification Penile Tissue
Gel Running Conditions Reduced Denaturing (4-20% mini gel)
Blocking step SuperBlock Blocking Buffer (PBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Miss. gwen Lagoda

Verified customer

提交于 Jul 26 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Epithelial, neutrophil, monocyte)
Loading amount 500000 cells
Specification Epithelial, neutrophil, monocyte
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris gel)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

提交于 Sep 09 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Rat Tissue lysate - whole (Penile Tissue)
Loading amount 80 µg
Specification Penile Tissue
Gel Running Conditions Reduced Denaturing (4-20% gel)
Blocking step Starting Block T20 (PBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Miss. gwen Lagoda

Verified customer

提交于 Jul 29 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"