Overview

  • Product name
    Anti-Ras antibody [EP1125Y]
    See all Ras primary antibodies
  • Description
    Rabbit monoclonal [EP1125Y] to Ras
  • Host species
    Rabbit
  • Specificity
    ab52939 is predicted to react with H-Ras, N-Ras and K-Ras.
  • Tested applications
    Suitable for: ICC/IF, WB, IP, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to Human Ras aa 1-100 (N terminal).

  • Positive control
    • Flow Cyt: PC-12 cells. WB: Jurkat, 293T, RAW 246.7, Neuro-2a, PC-12 and C6 lysates; Human Ras full length protein. ICC/IF: MCF7 cells. IP: Jurkat whole cell 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

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab52939 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
ICC/IF 1/500.
WB 1/5000. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).

For unpurified use at 1/10000- 1/50000.

IP 1/20.

For unpurified use at 1/30. 

Flow Cyt 1/30.

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

For unpurified use at 1/100. 

Target

  • Function
    Ras proteins bind GDP/GTP and possess intrinsic GTPase activity.
  • Involvement in disease
    Defects in HRAS are the cause of faciocutaneoskeletal syndrome (FCSS) [MIM:218040]. A rare condition characterized by prenatally increased growth, postnatal growth deficiency, mental retardation, distinctive facial appearance, cardiovascular abnormalities (typically pulmonic stenosis, hypertrophic cardiomyopathy and/or atrial tachycardia), tumor predisposition, skin and musculoskeletal abnormalities.
    Defects in HRAS are the cause of congenital myopathy with excess of muscle spindles (CMEMS) [MIM:218040]. CMEMS is a variant of Costello syndrome.
    Defects in HRAS may be a cause of susceptibility to Hurthle cell thyroid carcinoma (HCTC) [MIM:607464]. Hurthle cell thyroid carcinoma accounts for approximately 3% of all thyroid cancers. Although they are classified as variants of follicular neoplasms, they are more often multifocal and somewhat more aggressive and are less likely to take up iodine than are other follicular neoplasms.
    Note=Mutations which change positions 12, 13 or 61 activate the potential of HRAS to transform cultured cells and are implicated in a variety of human tumors.
    Defects in HRAS are a cause of susceptibility to bladder cancer (BLC) [MIM:109800]. A malignancy originating in tissues of the urinary bladder. It often presents with multiple tumors appearing at different times and at different sites in the bladder. Most bladder cancers are transitional cell carcinomas. They begin in cells that normally make up the inner lining of the bladder. Other types of bladder cancer include squamous cell carcinoma (cancer that begins in thin, flat cells) and adenocarcinoma (cancer that begins in cells that make and release mucus and other fluids). Bladder cancer is a complex disorder with both genetic and environmental influences.
    Note=Defects in HRAS are the cause of oral squamous cell carcinoma (OSCC).
  • Sequence similarities
    Belongs to the small GTPase superfamily. Ras family.
  • Post-translational
    modifications
    Palmitoylated by the ZDHHC9-GOLGA7 complex. A continuous cycle of de- and re-palmitoylation regulates rapid exchange between plasma membrane and Golgi.
    S-nitrosylated; critical for redox regulation. Important for stimulating guanine nucleotide exchange. No structural perturbation on nitrosylation.
  • Cellular localization
    Cell membrane. Golgi apparatus membrane. The active GTP-bound form is localized most strongly to membranes than the inactive GDP-bound form (By similarity). Shuttles between the plasma membrane and the Golgi apparatus.
  • Information by UniProt
  • Database links
  • Alternative names
    • C-BAS/HAS antibody
    • c-H-ras antibody
    • C-HA-RAS1 antibody
    • CTLO antibody
    • GTPase HRas antibody
    • GTPase KRas antibody
    • GTPase NRas antibody
    • H-Ras-1 antibody
    • H-RASIDX antibody
    • Ha-Ras antibody
    • HAMSV antibody
    • HRAS antibody
    • HRAS1 antibody
    • K RAS2A antibody
    • K RAS2B antibody
    • K RAS4A antibody
    • K RAS4B antibody
    • K-RAS antibody
    • KRAS antibody
    • KRAS1 antibody
    • KRAS2 antibody
    • N-RAS antibody
    • N-terminally processed antibody
    • NRAS antibody
    • NRAS1 antibody
    • p21ras antibody
    • RASH_HUMAN antibody
    • RASH1 antibody
    • RASK2 antibody
    • Transforming protein p21 antibody
    • v Ha ras Harvey rat sarcoma viral oncogene homolog antibody
    • v Ki ras2 Kirsten rat sarcoma viral oncogene homolog antibody
    • v ras neuroblastoma RAS viral oncogene homolog antibody
    see all

Images

  • All lanes : Anti-Ras antibody [EP1125Y] (ab52939) at 1/5000 dilution (purified)

    Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysates
    Lane 2 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysates
    Lane 3 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysates
    Lane 4 : Neuro-2a (mouse neuroblastoma cell line) whole cell lysates

    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: 21 kDa
    Observed band size: 21 kDa



    Blocking and diluting buffer: 5% NFDM/TBST

  • Immunocytochemistry analysis of MCF7 (human breast adenocarcinoma cell line) cells labeling Ras with Purified ab52939 at 1:500 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor ® 594) 1:200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor ® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • ab52939 (purified) at 1:20 dilution (2μg) immunoprecipitating Ras in Jurkat whole cell lysate.

    Lane 1 (input): Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate 10μg
    Lane 2 (+): ab52939 & Jurkat whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab52939 in Jurkat whole cell lysate

    For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary antibody at 1:1000 dilution.
    Blocking and diluting buffer: 5% NFDM/TBST.

  • Flow Cytometry analysis of PC-12 (rat adrenal gland pheochromocytoma cell line) cells labeling Ras with purified ab52939 at 1:30 dilution (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluor ® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

  • Anti-Ras antibody [EP1125Y] (ab52939) at 1/5000 dilution (purified) + PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysates at 15 µg

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

    Predicted band size: 21 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

  • Anti-Ras antibody [EP1125Y] (ab52939) at 1/500000 dilution (unpurified) + C6 (rat glial tumor cell line) cell lysate at 10 µg/ml

    Secondary
    goat anti-rabbit HRP at 1/2000 dilution

    Predicted band size: 21 kDa
    Observed band size: 18 kDa
    why is the actual band size different from the predicted?

  • Unpurified ab52939 at 1/100 dilution staining Ras in permeabilized PC-12 (rat adrenal gland pheochromocytoma cell line) cells by flow cytometry (red). Rabbit IgG negative control (green).

  • Anti-Ras antibody [EP1125Y] (ab52939) at 1/500 dilution (unpurified) + Human Ras full length protein (ab56526) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 21 kDa


    Exposure time: 1 minute

References

This product has been referenced in:
  • Yang Z  et al. miR-143-3p regulates cell proliferation and apoptosis by targeting IGF1R and IGFBP5 and regulating the Ras/p38 MAPK signaling pathway in rheumatoid arthritis. Exp Ther Med 15:3781-3790 (2018). Read more (PubMed: 29581736) »
  • Cheng L  et al. Zoledronate dysregulates fatty acid metabolism in renal tubular epithelial cells to induce nephrotoxicity. Arch Toxicol 92:469-485 (2018). Read more (PubMed: 28871336) »
See all 28 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Cancer Cell lines)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
25 µg
Specification
Cancer Cell lines
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Dec 21 2017

Application
Western blot
Sample
Human Cell lysate - nuclear (Vascular smooth muscle cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
Vascular smooth muscle cells
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Nov 21 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (Arachnoidal cells)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
15 µg
Specification
Arachnoidal cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Mar 02 2016

Answer

Thank you for kindly confirming these details. I am sorry these antibodies have not worked for this customer.

As requested, we are pleased to arrange the alternative free of charge replacements. I can confirm that 1 vial of ab87980 and 1 vial of ab6992 have been added to your order number PO NL-284-Dated:-07-04-2012. This has Abcam order reference 1067372,

I would like to reassure you that the free of charge replacement vials are also covered by our Abpromise guarantee. Should the customer still be experiencing difficulties with the new vial, or if you have any further questions, please do not hesitate to let me know.

Thank you for your help and cooperation with this case. Please do not hesitate to contact me if you need anything further.

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Answer

Thank you for taking the time to complete our questionnaire and contact us. I am sorry to hear the customer hashad difficulty obtaining satisfactory results from the free of charge replacement vials.

The details you have kindly provided will provide us with vital information for our monitoring of product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Reviewing the details, I am sorry there very few tips to provide on this occasion to help improve the results.

As there antibodies are regrettably not working for the customer, I can suggest an appropriatecourse of action on thisoccasion would be to provide acredit note in compensation for the original 2 vials. I would be pleased to arrange this for you.

Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (A549)
Loading amount
50 µg
Specification
A549
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Sep 17 2010

Answer

Thank you for your enquiry. I have obtained some further information on the flow cytometry protocol used to test this antibody. 2% paraformaldehyde was used for fixation: Solutions and Reagents: 1. 1X PBS 2. Blocking buffer 0.5% BSA in 1X PBS 3. 2% paraformaldehyde (1% solution – optional for storing samples) 4. 1X FACS permeabilisation solution 5. Fluorescently conjugated secondary antibody Protocol: 1. Collect 1x106 cells/sample 2. Wash cells once in blocking buffer 3. Fix cells with 2% paraformaldehyde and incubate at RT for 10 mins 4. Wash cells once with blocking buffer 5. Add 0.5ml 1X FACS permeabilising solution and incubate at RT for 10 mins 6. Wash cells once with blocking buffer 7. Incubate cells in blocking buffer for 30 min at RT 8. Add primary antibody at appropriate dilution and incubate for 30 mins RT. 9. Wash twice with blocking buffer and incubate with fluorescently – conjugated secondary antibody for 30 min RT 10. Wash cells twice with blocking buffer 11. Re-suspend cells in 1X PBS and analyse on flow cytometer. Samples can be kept in 1% paraformaldehyde at 4oC overnight. I hope this is helpful. Should you have any further questions, please do not hesitate to contact us again.

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