For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome

Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
United States
Your country/region is currently set to:

If incorrect, please enter your country/region into the box below, to view site information related to your country/region.

Call (888) 77-ABCAM (22226) or contact us
Need help? Contact us

  • My account
  • Sign out
Sign in or Register with us

Welcome

Sign in or

Don't have an account?

Register with us
My basket
Quick order
Abcam homepage

  • Research Products
    By product type
    Primary antibodies
    Secondary antibodies
    ELISA and Matched Antibody Pair Kits
    Cell and tissue imaging tools
    Cellular and biochemical assays
    Proteins and Peptides
    By product type
    Proteomics tools
    Agonists, activators, antagonists and inhibitors
    Cell lines and Lysates
    Multiplex miRNA assays
    Multiplex Assays
    By research area
    Cancer
    Cardiovascular
    Cell Biology
    Epigenetics
    Metabolism
    Developmental Biology
    By research area
    Immunology
    Microbiology
    Neuroscience
    Signal Transduction
    Stem Cells
  • Customized Products & Partnerships
    Customized Products & Partnerships

    Customized products and commercial partnerships to accelerate your diagnostic and therapeutic programs.

    Customized products

    Partner with us

  • Support
    Support hub

    Access advice and support for any research roadblock

    View support hub

    Protocols

    Your experiments laid out step by step

    View protocols

  • Events
    • Conference calendar
    • Cancer
    • Cardiovascular
    • Epigenetics & Nuclear signaling
    • Immunology
    • Neuroscience
    • Stem cells
    • Tradeshows
    • Scientific webinars
    Keep up to date with the latest events

    Full event breakdown with abstracts, speakers, registration and more

    View global event calendar

  • Pathways
    Cell signalling pathways

    View all pathways

    View all interactive pathways

  1. Link

    products/primary-antibodies/grb2-antibody-y301-ab32111.pdf

  1. Send me a copy of this email
    I agree to the terms and conditions.
Signal Transduction Adapters Cytoplasmic
Share by email
Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-GRB2 antibody [Y301] (ab32111)

  • Datasheet
  • SDS
Reviews (1)Q&A (2)References (4)

Product price, shipping and contact information

Currently unavailable

Sorry, we can't display this right now.

Please contact us to place your order, or try again later.

 

Loading size & price…

 

Shipping and order information

Shipping info

Promotion Information

Abpromise

Guaranteed product quality, expert customer support.

Find out more.

Western blot - Anti-GRB2 antibody [Y301] (ab32111)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRB2 antibody [Y301] (ab32111)
  • Immunocytochemistry/ Immunofluorescence - Anti-GRB2 antibody [Y301] (ab32111)
  • Flow Cytometry (Intracellular) - Anti-GRB2 antibody [Y301] (ab32111)
  • Western blot - Anti-GRB2 antibody [Y301] (ab32111)
  • Anti-GRB2 antibody [Y301] (ab32111)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y301] to GRB2
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

You may also be interested in

Protein
Product image
Recombinant Human GRB2 protein (ab116430)
Secondary
Product image
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Knockout
Product image
Human GRB2 knockout HCT116 cell line (ab273715)

View more associated products

Overview

  • Product name

    Anti-GRB2 antibody [Y301]
    See all GRB2 primary antibodies
  • Description

    Rabbit monoclonal [Y301] to GRB2
  • Host species

    Rabbit
  • Specificity

    ab32111 recognises GRB2. This antibody can also detect the splice isoform GRB3-3 of GRB2.
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)more details
    Unsuitable for: IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human GRB2 aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • Human breast carcinoma, PC12 cells WB: HCT116, HeLa, PC-12, NIH/3T3, HEK-293, RAW 264.7 and C6 cell lysates
  • 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    Y301
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Adapters
    • Cytoplasmic
    • Cancer
    • Signal transduction
    • G protein signaling
    • Small G proteins
    • Ras family

Associated products

  • Alternative Versions

    • Anti-GRB2 antibody [Y301] - BSA and Azide free (ab247231)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human GRB2 knockout HCT116 cell line (ab273715)
  • KO cell lysates

    • Human GRB2 knockout HCT116 cell lysate (ab275248)
  • Positive Controls

    • HeLa whole cell lysate (ab29545)
    • NIH/3T3 whole cell lysate (ab7179)
  • Recombinant Protein

    • Recombinant Human GRB2 protein (ab116430)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab32111 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)
1/1000 - 1/5000. Predicted molecular weight: 25 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100 - 1/250.
Flow Cyt (Intra)
1/40.

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

Notes
WB
1/1000 - 1/5000. Predicted molecular weight: 25 kDa.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
1/100 - 1/250.
Flow Cyt (Intra)
1/40.

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

Application notes
Is unsuitable for IP.

Target

  • Function

    Adapter protein that provides a critical link between cell surface growth factor receptors and the Ras signaling pathway.
    Isoform GRB3-3 does not bind to phosphorylated epidermal growth factor receptor (EGFR) but inhibits EGF-induced transactivation of a RAS-responsive element. Isoform GRB3-3 acts as a dominant negative protein over GRB2 and by suppressing proliferative signals, may trigger active programmed cell death.
  • Sequence similarities

    Belongs to the GRB2/sem-5/DRK family.
    Contains 1 SH2 domain.
    Contains 2 SH3 domains.
  • Domain

    The SH3 domains mediate interaction with RALGPS1 and SHB.
  • Cellular localization

    Golgi apparatus.
  • Target information above from: UniProt accession P62993 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2885 Human
    • Entrez Gene: 14784 Mouse
    • Entrez Gene: 81504 Rat
    • Omim: 108355 Human
    • SwissProt: P62993 Human
    • SwissProt: Q60631 Mouse
    • SwissProt: P62994 Rat
    • Unigene: 444356 Human
    • Unigene: 708119 Human
    • Unigene: 439649 Mouse
    • Unigene: 3360 Rat
    see all
  • Alternative names

    • Abundant SRC homology antibody
    • Adapter protein GRB2 antibody
    • ASH antibody
    • Ash protein antibody
    • EGFRBP GRB2 antibody
    • Epidermal growth factor receptor binding protein antibody
    • Epidermal growth factor receptor binding protein GRB2 antibody
    • GRB 2 antibody
    • GRB2 adapter protein antibody
    • Grb2 antibody
    • GRB2_HUMAN antibody
    • Grb3 3 antibody
    • Growth factor receptor bound protein 2 antibody
    • Growth factor receptor bound protein 3 antibody
    • Growth factor receptor-bound protein 2 antibody
    • HT027 antibody
    • MST084 antibody
    • MSTP084 antibody
    • NCKAP2 antibody
    • OTTHUMP00000166096 antibody
    • OTTHUMP00000166097 antibody
    • OTTHUMP00000166098 antibody
    • Protein ash antibody
    • SEM5 antibody
    • SH2/SH3 adapter GRB2 antibody
    see all

Images

  • Western blot - Anti-GRB2 antibody [Y301] (ab32111)
    Western blot - Anti-GRB2 antibody [Y301] (ab32111)
    All lanes : Anti-GRB2 antibody [Y301] (ab32111) at 1/2000 dilution

    Lane 1 : Wild-type HCT116 cell lysate
    Lane 2 : GRB2 knockout HCT116 cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : HEK293 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 25 kDa
    Observed band size: 25 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab32111 observed at 25 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab32111 was shown to react with GRB2 in HCT 116 wild-type cells in western blot with loss of signal observed in GRB2 knockout cell line ab273715 (GRB2 knockout cell lysate ab275248). HCT 116 wild-type and GRB2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab32111 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRB2 antibody [Y301] (ab32111)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRB2 antibody [Y301] (ab32111)

    Immunohistochemical analysis of GRB2 expression in paraffin embedded human breast carcinoma tissue section, using 1/100 ab32111. Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-GRB2 antibody [Y301] (ab32111)
    Immunocytochemistry/ Immunofluorescence - Anti-GRB2 antibody [Y301] (ab32111)
    Immunofluorescent analysis of GRB2 expression in PC12 cells, using 1/100 ab32111.
  • Flow Cytometry (Intracellular) - Anti-GRB2 antibody [Y301] (ab32111)
    Flow Cytometry (Intracellular) - Anti-GRB2 antibody [Y301] (ab32111)

    Overlay histogram showing SH-SY5Y cells stained with ab32111 (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 (ab32111, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

     

  • Western blot - Anti-GRB2 antibody [Y301] (ab32111)
    Western blot - Anti-GRB2 antibody [Y301] (ab32111)
    All lanes : Anti-GRB2 antibody [Y301] (ab32111) at 1/5000 dilution (purified)

    Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
    Lane 4 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate
    Lane 5 : RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
    Lane 6 : C6 (Rat glial tumor glial cell) whole cell lysate

    Lysates/proteins at 15 µg per lane.

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

    Predicted band size: 25 kDa
    Observed band size: 25 kDa



    Blocking/Diluting Buffer and concentration: 5% NFDM/TBST

    We are unsure how to define the extra bands.

  • Anti-GRB2 antibody [Y301] (ab32111)
    Anti-GRB2 antibody [Y301] (ab32111)

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 ab32111? Please let us know so that we can cite the reference in this datasheet.

ab32111 has been referenced in 4 publications.

  • Ren WM  et al. Neuroprotective Effects of ZiBuPiYin Recipe on db/db Mice via PI3K-Akt Signaling Pathway by Activating Grb2. Neural Plast 2021:8825698 (2021). PubMed: 33603781
  • Zhang Y  et al. Transcription factor PAX4 facilitates gastric cancer progression through interacting with miR-27b-3p/Grb2 axis. Aging (Albany NY) 13:16786-16803 (2021). PubMed: 34162761
  • Xiang YP  et al. Y772 phosphorylation of EphA2 is responsible for EphA2-dependent NPC nasopharyngeal carcinoma growth by Shp2/Erk-1/2 signaling pathway. Cell Death Dis 11:709 (2020). PubMed: 32848131
  • Parodi B  et al. Fumarates modulate microglia activation through a novel HCAR2 signaling pathway and rescue synaptic dysregulation in inflamed CNS. Acta Neuropathol 130:279-95 (2015). WB . PubMed: 25920452

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-3 of 3 Abreviews or Q&A

Western blot abreview for Anti-GRB2 antibody [Y301]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Raji)
Loading amount
40 µg
Specification
Raji
Gel Running Conditions
Reduced Denaturing (12%)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Read More
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 Sep 11 2009

Question

What is the recommended antigen retrieval buffer for these three antibodies?

Read More

Abcam community

Verified customer

Asked on Jan 23 2012

Answer

Thank you for contacting us.

The following standard heat-mediated citrate buffer antigen retrieval was used for the IHC validation of ab32111 and ab111031. Antibody ab111586 used citrate buffer in a pressure cooker at 120C for 5 minutes but a steamer or waterbath at 95-100C will be an acceptable substitute.

Place slides in a container of 0.01 M sodium citrate buffer, pH 6.0 heated to 100°C and maintain this temperature for 20 minutes by placing the container in a steamer or heated waterbath. After 20 minutes, remove from heat and let stand at room temperature in buffer for 20 minutes. Rinse in 1X TBS with Tween (TBST) for 1 minute at RT.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Read More

Abcam Scientific Support

Answered on Jan 23 2012

Question

What was the antigen retrieval method that was used to generate the picture on your datasheet?

Read More

Abcam community

Verified customer

Asked on Oct 24 2011

Answer

We used a heat mediated antigen retrieval method with sodium citrate, pH. 6.0.

Read More

Abcam Scientific Support

Answered on Oct 24 2011

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Get resources and offers direct to your inbox Sign up
A-Z by research area
  • Cancer
  • Cardiovascular
  • Cell biology
  • Developmental biology
  • Epigenetics & Nuclear signaling
  • Immunology
  • Metabolism
  • Microbiology
  • Neuroscience
  • Signal transduction
  • Stem cells
A-Z by product type
  • Primary antibodies
  • Secondary antibodies
  • Biochemicals
  • Isotype controls
  • Flow cytometry multi-color selector
  • Kits
  • Loading controls
  • Lysates
  • Peptides
  • Proteins
  • Slides
  • Tags and cell markers
  • Tools & Reagents
Help & support
  • Support
  • Make an Inquiry
  • Protocols & troubleshooting
  • Placing an order
  • RabMAb products
  • Biochemical product FAQs
  • Training
  • Browse by Target
Company
  • Corporate site
  • Investor relations
  • Company news
  • Careers
  • About us
  • Blog
Events
  • Tradeshows
  • Conferences
International websites
  • abcam.cn
  • abcam.co.jp

Join with us

  • LinkedIn
  • facebook
  • Twitter
  • YouTube
  • Terms of sale
  • Website terms of use
  • Cookie policy
  • Privacy policy
  • Legal
  • Modern slavery statement
© 1998-2023 Abcam plc. All rights reserved.