Overview

  • Product name
    Anti-RASA1 antibody [EP536Y]
    See all RASA1 primary antibodies
  • Description
    Rabbit monoclonal [EP536Y] to RASA1
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICCmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human RASA1 aa 250-350 (internal sequence). The exact sequence is proprietary. From near the SH3 domain.
    Database link: P20936

  • Positive control
    • WB: Rat brain tissue lysate.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab40677 in the following tested applications.

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

Application Abreviews Notes
WB 1/2000. Detects a band of approximately 140 kDa (predicted molecular weight: 96 kDa).
IHC-P 1/50.
ICC 1/250 - 1/500.
  • Application notes
    Is unsuitable for Flow Cyt or IP.
  • Target

    • Function
      Inhibitory regulator of the Ras-cyclic AMP pathway. Stimulates the GTPase of normal but not oncogenic Ras p21.
    • Tissue specificity
      In placental villi, detected only in the trophoblast layer (cytotrophoblast and syncytiotrophoblast). Not detected in stromal, endothelial or Hofbauer cells (at protein level).
    • Involvement in disease
      Note=Mutations in the SH2 domain of RASA seem to be oncogenic and cause basal cell carcinomas.
      Defects in RASA1 are the cause of capillary malformation-arteriovenous malformation (CMAVM) [MIM:608354]. CMAVM is a disorder characterized by atypical capillary malformations that are multiple, small, round to oval in shape and pinkish red in color. These capillary malformations are associated with either arteriovenous malformation, arteriovenous fistula, or Parkes Weber syndrome.
      Defects in RASA1 are a cause of Parkes Weber syndrome (PKWS) [MIM:608355]. PKWS is a disorder characterized by a cutaneous flush with underlying multiple micro-arteriovenous fistulas, in association with soft tissue and skeletal hypertrophy of the affected limb.
    • Sequence similarities
      Contains 1 C2 domain.
      Contains 1 PH domain.
      Contains 1 Ras-GAP domain.
      Contains 2 SH2 domains.
      Contains 1 SH3 domain.
    • Post-translational
      modifications
      The N-terminus is blocked.
    • Cellular localization
      Cytoplasm.
    • Information by UniProt
    • Database links
    • Alternative names
      • CM AVM antibody
      • CMAVM antibody
      • DKFZp434N071 antibody
      • GAP antibody
      • GTPase activating protein antibody
      • GTPase-activating protein antibody
      • OTTHUMP00000222390 antibody
      • OTTHUMP00000222391 antibody
      • OTTHUMP00000222392 antibody
      • OTTHUMP00000222393 antibody
      • p120GAP antibody
      • p120RASGAP antibody
      • PKWS antibody
      • Ras GTPase-activating protein 1 antibody
      • RAS p21 protein activator (GTPase activating protein) 1 antibody
      • Ras p21 protein activator antibody
      • RASA antibody
      • RASA1 antibody
      • RASA1_HUMAN antibody
      • RasGAP antibody
      • Triphosphatase activating protein antibody
      see all

    Images

    • Anti-RASA1 antibody [EP536Y] (ab40677) at 1/2000 dilution + Rat brain lysate at 10 µg

      Predicted band size: 96 kDa
      Observed band size: 140 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 55 kDa (possible cleavage fragment)

    • Immunohistochemical analysis of paraffin embedded human ovary carcinoma using ab40677 diluted 1:50.

    References

    This product has been referenced in:
    • Huang J  et al. Co-expression and significance of Dok2 and Ras p21 protein activator 1 in breast cancer. Oncol Lett 14:5386-5392 (2017). Read more (PubMed: 29098030) »
    • Teng HW  et al. Protein tyrosine phosphatase 1B targets PITX1/p120RasGAP thus showing therapeutic potential in colorectal carcinoma. Sci Rep 6:35308 (2016). Read more (PubMed: 27752061) »
    See all 4 Publications for this product

    Customer reviews and Q&As

    Answer

    Thank you for your enquiry.

    I can confirm the ICC testing protocol for which I have copied below. I hope this will be helpful. Please note, this would be a guideline only and may require some individual optimization.

    With regards to flow cytometry, this antibody has not yet been tested in this application. On our datasheets, we list the applications and species which have been tested and that we can provide our guarantee for. If we have data and information to indicate an antibody is not suitable for a particular application or species, we will also note this on the datasheet. Therefore, I would suggest this antibody is quite likely to work in flow cytometry, however we are not able to guarantee this without further testing. If you would like to try the antibody in this application, please do not hesitate to let me know before you purchase as you may be eligible for our testing discount program.

    I hope this information will be useful. If you have any further questions, please do not hesitate to contact me.

    ICC protocol:

    1. Solutions and reagents

    1.1. Washing buffer:

    PBST washing buffer: 1X PBS / 0.1% Tween-20 (Dulbecco’s Phosphate Buffered Saline)

    1.2. 2% Paraformaldehyde (prepare fresh by dissolving paraformaldehyde in 1X PBS by heating

    at 70oC until dissolving. Use cold.)

    1.3. 0.1% Triton X-100

    1.4. Blocking buffer:

    PBS (Dulbecco's Phosphate Buffered Saline) + 10% serum (serum origin depends on the host of the secondary antibody)

    1.5. Mounting medium for fluorescence.

    1.6 Acetone (Production Grade)

    2. Protocol

    2.1. Grow cells in chamber slides or in 6-well tissue plates containing sterile coverslips. Prior to fixation, cells should not reach more than 80% confluency.

    2.2. Fixation

    2.2.1. Remove medium from chamber slides or 6-well plates. Wash once with PBS-T.

    2.2.2. Fix cells with 2% paraformaldehyde for 20 min. For HeLa cells only, fix with acetone

    at -20oC for 5 mintues and continue to step 2.2.5.

    2.2.3. Wash cells twice with 1XPBST.

    2.2.4. Permeabilize cells with 0.1% Triton-X100 for 5 min

    2.2.5. Wash cells twice with 1XPBST.

    2.3. Blocking

    2.3.1. Block cells with blocking buffer for 1 hour at room temperature (option - O/N 4oC).

    2.4. Staining

    2.4.1. Dilute primary antibody in the blocking buffer per recommendation on the data sheet.

    2.4.2. Apply primary antibody on the cells and incubate overnight at 4oC.

    2.4.3. Wash cells twice with 1X PBST.

    2.4.4. Incubate cells in a dilution of fluorescently-labeled secondary antibody in PBS for 45 min at room temperature in the dark.

    2.4.5. Wash cells three times with 1X PBST.

    2.4.6. Counterstain cells with DAPI in a concentration of 300 nM in PBS for 5 min in the dark.

    2.4.7. Wash cells three times with 1X PBST.

    2.4.8. Mount slides with medium for fluorescent staining.

    2.4.9. Store slides in the dark

    Read More

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

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