Recombinant 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
Related conjugates and formulations
Overview
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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.
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Positive control
- Human breast carcinoma, PC12 cells WB: HCT116, HeLa, PC-12, NIH/3T3, HEK-293, RAW 264.7 and C6 cell lysates
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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
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
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y301 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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 |
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WB | (1) |
1/1000 - 1/5000. Predicted molecular weight: 25 kDa.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF |
1/100 - 1/250.
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Flow Cyt (Intra) |
1/40.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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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. |
Target
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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. - Information by UniProt
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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
see all -
Alternative names
- Abundant SRC homology antibody
- Adapter protein GRB2 antibody
- ASH antibody
see all
Images
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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 kDaLanes 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.
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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.
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Immunofluorescent analysis of GRB2 expression in PC12 cells, using 1/100 ab32111.
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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.
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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 kDaBlocking/Diluting Buffer and concentration: 5% NFDM/TBST
We are unsure how to define the extra bands.
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.
Datasheets and documents
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SDS download
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Datasheet download
References (4)
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