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Signal Transduction Second Messenger Nucleotide Messenger GTP
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RecombinantRabMAb

Recombinant Anti-RAP1GAP antibody [Y134] (ab32373)

  • Datasheet
  • SDS
Reviews (1)Q&A (1)References (4)

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Western blot - Anti-RAP1GAP antibody [Y134] (ab32373)
  • Western blot - Anti-RAP1GAP antibody [Y134] (ab32373)
  • Western blot - Anti-RAP1GAP antibody [Y134] (ab32373)
  • Western blot - Anti-RAP1GAP antibody [Y134] (ab32373)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAP1GAP antibody [Y134] (ab32373)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAP1GAP antibody [Y134] (ab32373)
  • Immunocytochemistry/ Immunofluorescence - Anti-RAP1GAP antibody [Y134] (ab32373)
  • Immunocytochemistry/ Immunofluorescence - Anti-RAP1GAP antibody [Y134] (ab32373)
  • Flow Cytometry - Anti-RAP1GAP antibody [Y134] (ab32373)
  • Flow Cytometry - Anti-RAP1GAP antibody [Y134] (ab32373)
  • Immunoprecipitation - Anti-RAP1GAP antibody [Y134] (ab32373)
  • Anti-RAP1GAP antibody [Y134] (ab32373)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y134] to RAP1GAP
  • Suitable for: ICC/IF, WB, IHC-P, Flow Cyt, IP
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-RAP1GAP antibody [Y134]
    See all RAP1GAP primary antibodies
  • Description

    Rabbit monoclonal [Y134] to RAP1GAP
  • Host species

    Rabbit
  • Specificity

    ab32373 recognises Rap1 GTPase-activating protein 1
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Rat
    WB
    Mouse
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide within Human RAP1GAP aa 1-100 (N terminal). The exact sequence is proprietary.
    (Peptide available as ab139973)

  • Positive control

    • WB: Jurkat, SH-SY5Y cell lysate, mouse and rat brain lysate IHC: Human breast cancer ICC/IF: HeLa cells IP: C6 cell lysate
  • General notes

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    Y134
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Second Messenger
    • Nucleotide Messenger
    • GTP
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Cancer susceptibility
    • Tumor Suppressors
    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • Other

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-RAP1GAP antibody [Y134] (ab202756)
    • Alexa Fluor® 647 Anti-RAP1GAP antibody [Y134] (ab202758)
    • Alexa Fluor® 555 Anti-RAP1GAP antibody [Y134] (ab203452)
    • Anti-RAP1GAP antibody [Y134] - BSA and Azide free (ab247250)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Immunizing Peptide (Blocking)

    • RAP1GAP peptide (ab139973)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • Jurkat whole cell lysate (ab7899)

Applications

The Abpromise guarantee

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.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Human
ICC/IF
Human
IHC-P
Human
IP
Rat
WB
Mouse
Human
Application Abreviews Notes
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)
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.
Notes
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. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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.

Target

  • Function

    GTPase activator for the nuclear Ras-related regulatory protein RAP-1A (KREV-1), converting it to the putatively inactive GDP-bound state.
  • Tissue specificity

    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.
  • Sequence similarities

    Contains 1 GoLoco domain.
    Contains 1 Rap-GAP domain.
  • Cellular localization

    Golgi apparatus membrane.
  • Target information above from: UniProt accession P47736 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 5909 Human
    • Entrez Gene: 110351 Mouse
    • Entrez Gene: 313644 Rat
    • Omim: 600278 Human
    • SwissProt: P47736 Human
    • SwissProt: A2ALS5 Mouse
    • Unigene: 148178 Human
    • Unigene: 180763 Mouse
    • Unigene: 18036 Rat
    see all
  • Alternative names

    • 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

Images

  • Western blot - Anti-RAP1GAP antibody [Y134] (ab32373)
    Western blot - Anti-RAP1GAP antibody [Y134] (ab32373)
    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
    All lanes : 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

  • Western blot - Anti-RAP1GAP antibody [Y134] (ab32373)
    Western blot - Anti-RAP1GAP antibody [Y134] (ab32373)
    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

  • Western blot - Anti-RAP1GAP antibody [Y134] (ab32373)
    Western blot - Anti-RAP1GAP antibody [Y134] (ab32373)
    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

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

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAP1GAP antibody [Y134] (ab32373)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAP1GAP antibody [Y134] (ab32373)
    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.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAP1GAP antibody [Y134] (ab32373)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RAP1GAP antibody [Y134] (ab32373)

    Immunohistochemical staining of paraffin-embedded human breast cancer tissue using unpurified ab32373 at 1/100 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-RAP1GAP antibody [Y134] (ab32373)
    Immunocytochemistry/ Immunofluorescence - Anti-RAP1GAP antibody [Y134] (ab32373)

    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-RAP1GAP antibody [Y134] (ab32373)
    Immunocytochemistry/ Immunofluorescence - Anti-RAP1GAP antibody [Y134] (ab32373)

    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.

  • Flow Cytometry - Anti-RAP1GAP antibody [Y134] (ab32373)
    Flow Cytometry - Anti-RAP1GAP antibody [Y134] (ab32373)
    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).
  • Flow Cytometry - Anti-RAP1GAP antibody [Y134] (ab32373)
    Flow Cytometry - Anti-RAP1GAP antibody [Y134] (ab32373)

    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.

  • Immunoprecipitation - Anti-RAP1GAP antibody [Y134] (ab32373)
    Immunoprecipitation - Anti-RAP1GAP antibody [Y134] (ab32373)

    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 Detection Reagent (ab131366) was used for detection (1/1000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST

  • Anti-RAP1GAP antibody [Y134] (ab32373)
    Anti-RAP1GAP antibody [Y134] (ab32373)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (4)

Publishing research using ab32373? Please let us know so that we can cite the reference in this datasheet.

ab32373 has been referenced in 4 publications.

  • 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 . 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 . PubMed: 24642466
  • Hewer RC  et al. PKA and Epac synergistically inhibit smooth muscle cell proliferation. J Mol Cell Cardiol 50:87-98 (2011). WB ; Rat . PubMed: 20971121
  • McAvoy T  et al. Phosphorylation of Rap1GAP, a striatally enriched protein, by protein kinase A controls Rap1 activity and dendritic spine morphology. Proc Natl Acad Sci U S A 106:3531-6 (2009). PubMed: 19218462

Customer reviews and Q&As

Show All Reviews Q&A
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1-2 of 2 Abreviews or Q&A

Question

Is it possible to order the immunogen peptide for antibody ab32373 from you? We are using this antibody and need to proof its specificity!

Therefor we need the immunogen peptide to perform a competition assay as a negative control.

Read More

Abcam community

Verified customer

Asked on Sep 28 2012

Answer

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

https://www.abcam.com/RAP1GAP-peptide-ab139973.html (or use the following: https://www.abcam.com/RAP1GAP-peptide-ab139973.html).

I wish you good luck with your experiments and a good weekend.

Read More

Abcam Scientific Support

Answered on Sep 28 2012

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-RAP1GAP antibody [Y134]

Excellent
Abreviews
Abreviews
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
Read More

Abcam user community

Verified customer

Submitted Mar 07 2008

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