RabMAb

Anti-RAP1GAP抗体[Y134] (ab32373)

概述

  • 产品名称Anti-RAP1GAP抗体[Y134]
    参阅全部 RAP1GAP 一抗
  • 描述
    兔单克隆抗体[Y134] to RAP1GAP
  • 特异性ab32373 recognises Rap1 GTPase-activating protein 1
  • 经测试应用适用于: ICC/IF, WB, IHC-P, Flow Cyt, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human RAP1GAP aa 1-100 (N terminal).
    (Peptide available as ab139973)

  • 阳性对照
    • WB: Jurkat, SH-SY5Y cell lysate, mouse and rat brain lysate IHC: Human breast cancer ICC/IF: HeLa cells IP: C6 cell lysate
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

     

    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.

    Alternative versions available:

    Anti-RAP1GAP antibody (Alexa Fluor® 488) [Y134] (ab202756)
    Anti-RAP1GAP antibody (Alexa Fluor® 647) [Y134] (ab202758)
    Anti-RAP1GAP antibody (Alexa Fluor® 555) [Y134] (ab203452)

     

性能

应用

Our Abpromise guarantee covers the use of ab32373 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
ICC/IF 1/100 - 1/250.
WB 1/10000 - 1/50000. Detects a band of approximately 95 kDa (predicted molecular weight: 95 kDa).Can be blocked with RAP1GAP peptide (ab139973).
IHC-P 1/1000.
Flow Cyt 1/30 - 1/50.

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

IP 1/20 - 1/60.

靶标

  • 功能GTPase activator for the nuclear Ras-related regulatory protein RAP-1A (KREV-1), converting it to the putatively inactive GDP-bound state.
  • 组织特异性Significant expression seen in the brain, kidney and pancreas. Abundant in the cerebral cortex and expressed at much lower levels in the spinal cord. Not detected in the lymphoid tissues.
  • 序列相似性Contains 1 GoLoco domain.
    Contains 1 Rap-GAP domain.
  • 细胞定位Golgi apparatus membrane.
  • Information by UniProt
  • 数据库链接
  • 别名
    • KIAA0474 antibody
    • Rap1 GTPase activating protein 1 antibody
    • RAP1 GTPase activating protein antibody
    • Rap1 GTPase-activating protein 1 antibody
    • RAP1GA1 antibody
    • Rap1ga1 protein antibody
    • Rap1GAP antibody
    • Rap1GAP1 antibody
    • RAP1GAPII antibody
    • RAPGAP antibody
    • RPGP1_HUMAN antibody
    see all

Anti-RAP1GAP antibody [Y134] 图像

  • All lanes : Anti-RAP1GAP antibody [Y134] (ab32373) at 1/10000 dilution (purified)

    Lane 1 : Mouse brain lysate
    Lane 2 : Rat brain lysate

    Lysates/proteins at 20 µg per lane.

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

    Predicted band size : 95 kDa
    Observed band size : 95 kDa

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Anti-RAP1GAP antibody [Y134] (ab32373) at 1/10000 dilution (purified) + Jurkat cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size : 95 kDa
    Observed band size : 95 kDa

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Anti-RAP1GAP antibody [Y134] (ab32373) at 1/10000 dilution (purified) + SH-SY5Y cell lysate at 20 µg

    Secondary
    Anti-rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

    Predicted band size : 95 kDa
    Observed band size : 95 kDa

    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Anti-RAP1GAP antibody [Y134] (ab32373) at 1/50000 dilution (unpurified) + Jurkat cell lysate

    Predicted band size : 95 kDa
    Observed band size : 95 kDa
  • Immunohistochemical staining of paraffin embedded human cerebral cortex with purified ab32373 at a working dilution of 1/1000. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • Immunohistochemical staining of paraffin-embedded human breast cancer tissue using unpurified ab32373 at 1/100 dilution.

    Paraffin-embedded human breast cancer tissue
    ab32373 at 1/100 dilution
  • Immunofluorescence staining of HeLa cells with purified ab32373 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4 % PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab32373 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • ICC/IF image of unpurified ab32373 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab32373, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h.Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing Jurkat cells fixed in 4% PFA and stained with purified ab32373 at a dilution of 1/30 (red line). The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit at a dilution of 1/500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
  • Overlay histogram showing SH-SY5Y cells stained with unpurified ab32373 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab32373, 1/100) 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.

  • ab32373 (purified) at 1/20 immunoprecipitating RAP1GAP in 10 μg C6 cell lysate (Lanes 1 and 2, observed at 82-95 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730). For western blotting, HRP Veriblot for IP (ab131366) was used as the secondary antibody (1/1000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST

Anti-RAP1GAP antibody [Y134] (ab32373)参考文献

This product has been referenced in:
  • Morales Johansson H  et al. Whole-genome analysis of temporal gene expression during early transdifferentiation of human lung alveolar epithelial type 2 cells in vitro. PLoS One 9:e93413 (2014). WB ; Human . Read more (PubMed: 24690998) »
  • Potla U  et al. Podocyte-specific RAP1GAP expression contributes to focal segmental glomerulosclerosis-associated glomerular injury. J Clin Invest 124:1757-69 (2014). WB, IF ; Mouse, Human . Read more (PubMed: 24642466) »

See all 4 Publications for this product

Product Wall

Thank you for your inquiry.

I am happy to confirm that the blocking peptide for ab32373 is added to our catalog under the product number:

ab139973

http://www.abcam.com/index.html?datasheet=139973 (or use the following: htt...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (prostate)
Specification prostate
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH9 OR Sodium citrate pH 6.0
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Mar 07 2008

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