Anti-GRB2 antibody [Y301] Recombinant (ab32111)

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 Cytmore 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, NIH3T3 cells, PC12 cells, HeLa cells.
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.

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, 0.05% BSA
Concentration information loading...
Purity

IgG fraction
Clonality

Monoclonal
Clone number

Y301
Isotype

IgG
Research areas

Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- HeLa whole cell lysate (ab29545)
- NIH 3T3 whole cell lysate (ab7179)

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		1/1000 - 1/2000. Predicted molecular weight: 25 kDa.
IHC-P		Use at an assay dependent concentration.
ICC/IF		1/100 - 1/250.
Flow Cyt		1/40. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Application notes

Is unsuitable for IP.

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

Western blot - Anti-GRB2 antibody [Y301] (ab32111)

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

Lane 1 : NIH 3T3 cell lysate
Lane 2 : PC12 cell lysate
Lane 3 : HeLa cell lysate

Predicted band size: 25 kDa
Observed band size: 25 kDa
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.
Immunocytochemistry/ Immunofluorescence - Anti-GRB2 antibody [Y301] (ab32111)

Immunofluorescent analysis of GRB2 expression in PC12 cells, using 1/100 ab32111.
Flow Cytometry - 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 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.

Datasheet

This product has been referenced in:

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 . Read more (PubMed: 25920452) »

See 1 Publication for this product

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

Customer reviews and Q&As

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1-3 of 3 Abreviews or Q&A

Question

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

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.

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?

Abcam community

Verified customer

Asked on Oct 24 2011

Answer

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

Abcam Scientific Support

Answered on Oct 24 2011

Abcam guarantees this product to work in the species/application used in this Abreview.

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

Abcam user community

Verified customer

Submitted Sep 11 2009

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

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Anti-GRB2 antibody [Y301] (ab32111)

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References

This product has been referenced in:

Customer reviews and Q&As

Western blot abreview for Anti-GRB2 antibody [Y301]