Key features and details
- Mouse monoclonal [2G9] to HEF1/NEDD-9
- Suitable for: WB
- Reacts with: Human
- Isotype: IgG1
Product nameAnti-HEF1/NEDD-9 antibody [2G9]
See all HEF1/NEDD-9 primary antibodies
DescriptionMouse monoclonal [2G9] to HEF1/NEDD-9
Tested applicationsSuitable for: WBmore details
Species reactivityReacts with: Human
Synthetic peptide corresponding to Human HEF1/NEDD-9 aa 50-400. Anti-HEF1 monoclonal antibody was produced by repeated immunizations with a synthetic peptide corresponding to amino acid residues 82-398 of human HEF1 protein (hHEF1, 843 aa, predicted MW 92.8 kDa)
Database link: Q14511
- WB: MCF7 lysate
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 0.424% Potassium phosphate, 0.87% Sodium chloride
Concentration information loading...
PurityProtein A purified
Light chain typekappa
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab288750 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000 - 1/5000. Predicted molecular weight: 92 kDa.
1/1000 - 1/5000. Predicted molecular weight: 92 kDa.
FunctionDocking protein which plays a central coordinating role for tyrosine-kinase-based signaling related to cell adhesion. May function in transmitting growth control signals between focal adhesions at the cell periphery and the mitotic spindle in response to adhesion or growth factor signals initiating cell proliferation. May play an important role in integrin beta-1 or B cell antigen receptor (BCR) mediated signaling in B- and T-cells. Integrin beta-1 stimulation leads to recruitment of various proteins including CRK, NCK and SHPTP2 to the tyrosine phosphorylated form.
Tissue specificityWidely expressed. Higher levels detected in kidney, lung, and placenta. Also detected in T-cells, B-cells and diverse cell lines. The protein has been detected in lymphocytes, in diverse cell lines, and in lung tissues.
Sequence similaritiesBelongs to the CAS family.
Contains 1 SH3 domain.
DomainContains a central domain containing multiple potential SH2-binding sites and a C-terminal domain containing a divergent helix-loop-helix (HLH) motif. The SH2-binding sites putatively bind CRK, NCK and ABL SH2 domains. The HLH motif confers specific interaction with the HLH proteins ID2, E12 and E47. It is absolutely required for the induction of pseudohyphal growth in yeast and mediates homodimerization and heterodimerization with p130cas.
The SH3 domain interacts with two proline-rich regions of focal adhesion kinase.
modificationsCell cycle-regulated processing produces four isoforms: p115, p105, p65, and p55. Isoform p115 arises from p105 phosphorylation and appears later in the cell cycle. Isoform p55 arises from p105 as a result of cleavage at a caspase cleavage-related site and it appears specifically at mitosis. The p65 isoform is poorly detected.
Focal adhesion kinase 1 phosphorylates the protein at the YDYVHL motif (conserved among all cas proteins). The SRC family kinases (FYN, SRC, LCK and CRK) are recruited to the phosphorylated sites and can phosphorylate other tyrosine residues. Ligation of either integrin beta-1 or B-cell antigen receptor on tonsillar B-cells and B-cell lines promotes tyrosine phosphorylation and both integrin and BCR-mediated tyrosine phosphorylation requires an intact actin network. In fibroblasts transformation with oncogene v-ABL results in an increase in tyrosine phosphorylation. Transiently phosphorylated following CD3 cross-linking and this phosphorylated form binds to CRK and C3G. A mutant lacking the SH3 domain is phosphorylated upon CD3 cross-linking but not upon integrin beta-1 cross-linking. Tyrosine phosphorylation occurs upon stimulation of the G-protein coupled C1a calcitonin receptor in rabbit. Calcitonin-stimulated tyrosine phosphorylation is mediated by calcium- and protein kinase C-dependent mechanisms and requires the integrity of the actin cytoskeleton.
Cellular localizationCytoplasm > cytoskeleton > spindle and Cytoplasm > cell cortex. Nucleus. Golgi apparatus. Cell projection > lamellipodium. Cytoplasm. Cell junction > focal adhesion. Localizes to both the cell nucleus and the cell periphery and is differently localized in fibroblasts and epithelial cells. In fibroblasts is predominantly nuclear and in some cells is present in the Golgi apparatus. In epithelial cells localized predominantly in the cell periphery with particular concentration in lamellipodia but is also found in the nucleus. Isoforms p105 and p115 are predominantly cytoplasmic and associate with focal adhesions while p55 associates with mitotic spindle.
- Information by UniProt
- Cas like docking antibody
- Cas scaffolding protein family member 2 antibody
- CAS-L antibody
Anti-HEF1/NEDD-9 antibody [2G9] (ab288750) at 1/1000 dilution + MCF7 lysate
IRDye®800 conjugated Sh-a-Mouse IgG [H&L] at 1/5000 dilution
Predicted band size: 92 kDa
Western blot using ab288750 monoclonal anti-HEF1 antibody (clone 2G9) antibody shows detection of a ~92 kDa band corresponding to HEF1 in MCF7 lysate [arrowhead]. Approximately 35 µg of lysate was loaded for SDS-PAGE followed by transfer onto nitrocellulose and reaction with a 1:1,000 dilution of anti-HEF1 antibody. Detection occurred using a 1:5,000 dilution of IRDye®800 conjugated Sh-a-Mouse IgG [H&L] for 45 min at room temperature (800 nm channel, green). Molecular weight estimation was made by comparison to prestained MW markers (indicated at left). IRDye®800 fluorescence image was captured using the Odyssey® Infrared Imaging System developed by LI-COR. IRDye is a trademark of LI-COR, Inc. Other detection systems will yield similar results.
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.
ab288750 has not yet been referenced specifically in any publications.