Name: Anti-GTPase HRAS antibody [Y132]
Brand: RabMAb
SKU: ab32417
Rating: 3 (6 reviews)

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Western blot - Anti-GTPase HRAS antibody [Y132] (ab32417)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [Y132] to GTPase HRAS
Suitable for: WB, IP
Knockout validated
Reacts with: Mouse, Rat, Human

Carrier Free

Product name

Anti-GTPase HRAS antibody [Y132]
See all GTPase HRAS primary antibodies
Description

Rabbit monoclonal [Y132] to GTPase HRAS
Host species

Rabbit
Specificity

Reactivity with other RAS members has not been tested.
Tested applications

Suitable for: WB, IPmore details
Unsuitable for: IHC
Species reactivity

Reacts with: Mouse, Rat, Human
Predicted to work with: Chicken
Immunogen

Synthetic peptide within Human GTPase HRAS aa 150 to the C-terminus (C terminal). The exact sequence is proprietary.
Positive control
- MCF7 and PC12 cell lysates and MCF7 cells.
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.

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

Y132
Isotype

IgG
Research areas

Alternative Versions
- Anti-GTPase HRAS antibody [Y132] - BSA and Azide free (ab239815)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human HRAS knockout HEK-293 cell line (ab260858)
KO cell lysates
- Human HRAS knockout HEK-293 cell lysate (ab261650)
Recombinant Protein
- Recombinant Human GTPase HRAS protein (ab93949)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32417 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	(6)	1/500 - 1/1000. Detects a band of approximately 21 kDa.
IP		1/50 - 1/60.

Notes
WB 1/500 - 1/1000. Detects a band of approximately 21 kDa.
IP 1/50 - 1/60.

Application notes

Is unsuitable for IHC.

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.
Target information above from: UniProt accession P01112 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 396229 Chicken
- Entrez Gene: 3265 Human
- Entrez Gene: 15461 Mouse
- Entrez Gene: 293621 Rat
- Omim: 190020 Human
- SwissProt: P08642 Chicken
- SwissProt: P01112 Human
- SwissProt: Q61411 Mouse
- SwissProt: P20171 Rat
- Unigene: 37003 Human
- Unigene: 334313 Mouse
- Unigene: 102180 Rat
see all
Alternative names
- C BAS/HAS antibody
- c H ras antibody
- C HA RAS1 antibody
- c has/bas p21 protein antibody
- c ras Ki 2 activated oncogene antibody
- c-H-ras antibody
- CTLO antibody
- GTP and GDP binding peptide B antibody
- GTPase HRas, N-terminally processed antibody
- H Ras 1 antibody
- H RASIDX antibody
- H-Ras-1 antibody
- Ha Ras antibody
- Ha Ras1 proto oncoprotein antibody
- Ha-Ras antibody
- HAMSV antibody
- Harvey rat sarcoma viral oncogene homolog antibody
- Harvey rat sarcoma viral oncoprotein antibody
- HRAS antibody
- HRAS1 antibody
- K ras antibody
- N ras antibody
- p19 H RasIDX protein antibody
- p21ras antibody
- Ras family small GTP binding protein H Ras antibody
- RASH_HUMAN antibody
- RASH1 antibody
- Transformation gene oncogene HAMSV antibody
- Transforming protein p21 antibody
- v Ha ras Harvey rat sarcoma viral oncogene homolog antibody
- VH Ras antibody
- vHa RAS antibody
see all

Western blot - Anti-GTPase HRAS antibody [Y132] (ab32417)

All lanes : Anti-GTPase HRAS antibody [Y132] (ab32417) at 1/500 dilution

Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : HRAS knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Lanes 1 - 4: Merged signal (red and green). Green - ab32417 observed at 21 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab32417 was shown to recognize HRAS in wild-type HEK-293 cells as signal was lost at the expected MW in HRAS knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and HRAS knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab32417 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-GTPase HRAS antibody [Y132] (ab32417)

All lanes : Anti-GTPase HRAS antibody [Y132] (ab32417) at 1/2500 dilution (purified)

Lane 1 : mouse brain lysate
Lane 2 : rat brain lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

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

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Western blot - Anti-GTPase HRAS antibody [Y132] (ab32417)

All lanes : Anti-GTPase HRAS antibody [Y132] (ab32417) at 1/1000 dilution (purified)

Lane 1 : MCF7 cell lysate
Lane 2 : HeLa cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

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

Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
Immunoprecipitation - Anti-GTPase HRAS antibody [Y132] (ab32417)

ab32417 (purified) at 1/60 immunoprecipitating GTPase in 10 μg mouse brain whole cell lysate (Lanes 1 and 2, observed at 21 kDa). Lane 3 - PBS. For western blotting, HRP Veriblot for IP Detection Reagent (ab131366) was used for detection (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
Western blot - Anti-GTPase HRAS antibody [Y132] (ab32417)

All lanes : Anti-GTPase HRAS antibody [Y132] (ab32417) at 1/500 dilution (unpurified)

Lane 1 : MCF7 cell lysate
Lane 2 : PC12 cell lysate

Observed band size: 21 kDa why is the actual band size different from the predicted?
Anti-GTPase HRAS antibody [Y132] (ab32417)

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

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab32417? Please let us know so that we can cite the reference in this datasheet.

ab32417 has been referenced in 15 publications.

Liu Z et al. CircZNF609 promotes cell proliferation, migration, invasion, and glycolysis in nasopharyngeal carcinoma through regulating HRAS via miR-338-3p. Mol Cell Biochem 476:175-186 (2021). PubMed: 32970285
Liu J et al. Ethanol Extract of the Infructescence of Platycarya strobilacea Sieb. et Zucc. Induces Methuosis of Human Nasopharyngeal Carcinoma Cells. Evid Based Complement Alternat Med 2020:2760979 (2020). PubMed: 32419796
Ong T et al. Siah2 integrates mitogenic and extracellular matrix signals linking neuronal progenitor ciliogenesis with germinal zone occupancy. Nat Commun 11:5312 (2020). PubMed: 33082319
Zhao T et al. DNA methylation-regulated QPCT promotes sunitinib resistance by increasing HRAS stability in renal cell carcinoma. Theranostics 9:6175-6190 (2019). PubMed: 31534544
Anta B et al. PGA1-induced apoptosis involves specific activation of H-Ras and N-Ras in cellular endomembranes. Cell Death Dis 7:e2311 (2016). WB . PubMed: 27468687

View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A

Submit a review Submit a question

1-9 of 9 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (Various cell lines)

Gel Running Conditions

Reduced Denaturing (4-15%)

Loading amount

25 µg

Specification

Various cell lines

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 04 2022

Application

Western blot

Sample

Mouse Cell lysate - whole cell (MEFs)

Gel Running Conditions

Reduced Denaturing (4-15%)

Loading amount

25 µg

Specification

MEFs

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 04 2022

Application

Western blot

Sample

Rat Cell lysate - whole cell (3611-RF cells)

Gel Running Conditions

Reduced Denaturing (4-15%)

Loading amount

25 µg

Specification

3611-RF cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 04 2022

Application

Western blot

Sample

Dog Cell lysate - whole cell (MDCK cells)

Gel Running Conditions

Reduced Denaturing (4-15%)

Loading amount

25 µg

Specification

MDCK cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 04 2022

Application

Western blot

Sample

Monkey Cell lysate - whole cell (COS-1 cells)

Gel Running Conditions

Reduced Denaturing (4-15%)

Loading amount

25 µg

Specification

COS-1 cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 04 2022

Application

Western blot

Sample

Mouse Cell lysate - whole cell (Colon cancer cells)

Loading amount

100 µg

Specification

Colon cancer cells

Gel Running Conditions

Reduced Denaturing

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 17 2011

Question

Product code: 96548
Inquiry: Is any of the following antibodies sold by you (ab96548, ab32417, ab16881, ab16907) corresponding to either sc29 or sc-520 from Santa Cruz? Many thanks,

Abcam community

Verified customer

Asked on Feb 20 2013

Answer

Thank you for contacting us.
ab97488 and ab6548 are alternative to sc-520. Please note this assumption is based on the location of immunogen only. We might not have similar species or application data.
ab140962 and ab140377 are expected alternatives for sc29.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Abcam Scientific Support

Answered on Feb 20 2013

Question

Dear technical support agent,
I will be happy to test the antibody and write an abreview. I will order the Ab tomorrow morning. Just let me know what I need to do next.
Many thanks,

Abcam community

Verified customer

Asked on Feb 15 2012

Answer

Thank you for your reply. I have created the discount code below, which can be redeemed for a free primary antibody after submitting a review of ab96548 and following the instructions below. Please let me know if you have any questions or if there is anything else that I can help you with.
DISCOUNT CODE: *** Expiration date: June 16, 2012
This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for ab96548 with mouse samples and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.
The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.
Any feedback that you can provide will be greatly appreciated, whether positive or negative. We look forward to receiving your Abreview and wish you luck with your research.
The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Abcam Scientific Support

Answered on Feb 15 2012

Question

I am looking for an antibody that is specific for murine H-Ras (no cross reactivity with K- or N-Ras) to be used for WB and/or IHC. Do you have anything interesting? I have already tried your ab32417 and I know that ab16881 cross reacts with N and Kras. What about ab64705 and ab96548? are they suitable for this use? could I have a trial size to test them?
Many thanks,

Abcam community

Verified customer

Asked on Feb 14 2012

Answer

Thank you for contacting us with your enquiry.
I have run some alignments between the immunogens of ab64705 and ab96548, and ab96548 is more specific for HRas. The immunogen of ab64705 overlaps quite a bit with N and KRas, however ab96548 shows no significant similarity to KRas and very little (47%) to NRas.
We only have the full size vials of all of our products and are not able to send trial sizes. However, as ab96548 has not been tested in murine samples, you are eligible for our testing discount program if you would like to try this antibody in mouse. For this program, you would initially purchase ab96548, test it in mouse, then submit an Abreview with your data. In exchange we will give you a discount code valid for one free primary antibody on a future order.
I hope this information will be useful. Please let me know if you have any questions, or if you would like to participate in our testing discount program, and I will be happy to arrange that for you.

Abcam Scientific Support

Answered on Feb 14 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

KO cell lines

KO cell lysates

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (15)

Customer reviews and Q&As

Western blot abreview for Anti-GTPase HRAS antibody [Y132]

Western blot abreview for Anti-GTPase HRAS antibody [Y132]

Western blot abreview for Anti-GTPase HRAS antibody [Y132]

Western blot abreview for Anti-GTPase HRAS antibody [Y132]

Western blot abreview for Anti-GTPase HRAS antibody [Y132]

Western blot abreview for Anti-GTPase HRAS antibody [Y132]

Post-translational
modifications