Overview

  • Product nameAnti-HEF1 antibody
    See all HEF1 primary antibodies
  • Description
    Rabbit polyclonal to HEF1
  • Tested applicationsSuitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 300 - 400 of Human HEF1.

    (Peptide available as ab39117.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa (Human epithelial carcinoma cell line) Jurkat (Human T cell lymphoblast-like cell line)

Properties

Applications

Our Abpromise guarantee covers the use of ab37161 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 1/250. Detects a band of approximately 120 kDa (predicted molecular weight: 110 kDa).
ICC/IF Use a concentration of 5 µg/ml.

Target

  • 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.
  • Post-translational
    modifications
    Cell 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
  • Database links
  • Alternative names
    • Cas like docking antibody
    • Cas scaffolding protein family member 2 antibody
    • CAS-L antibody
    • CAS2 antibody
    • CasL antibody
    • CASL_HUMAN antibody
    • CASS2 antibody
    • Crk associated substrate related antibody
    • Crk associated substrate related protein antibody
    • CRK-associated substrate-related protein antibody
    • dJ49G10.2 (Enhancer of Filamentation 1 (HEF1)) antibody
    • dJ49G10.2 antibody
    • dJ761I2.1 (enhancer of filamentation (HEF1)) antibody
    • dJ761I2.1 antibody
    • Enhancer of filamentation 1 antibody
    • Enhancer of filamentation 1 p55 antibody
    • HEF 1 antibody
    • HEF1 antibody
    • NEDD-9 antibody
    • Nedd9 antibody
    • Neural precursor cell expressed developmentally down-regulated protein 9 antibody
    • P105 antibody
    • Renal carcinoma antigen NY-REN-12 antibody
    see all

Anti-HEF1 antibody images

  • All lanes : Anti-HEF1 antibody (ab37161) at 1/250 dilution

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat whole cell lysate (ab7899)
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human HEF1 peptide (ab39117) at 1 µg/ml
    Lane 4 : Jurkat whole cell lysate (ab7899) with Human HEF1 peptide (ab39117) at 1 µg/ml

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 110 kDa
    Observed band size : 120 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 40 kDa. We are unsure as to the identity of these extra bands.
  • ab37161 staining HEF1 in Human glioblastoma cells by Immunocytochemistry/ Immunofluorescence. Cells were PFA-fixed and permeabilized in 0.1% Triton X-100 in PBS prior to blocking in 0.5% BSA in TBS-Tween for 20 minutes at room temperature. The primary antibody was diluted 1/50 in 0.5% BSA/PBS and incubated with the sample for 16 hours at 4°C. The secondary antibody was Cy3®-conjugated Goat anti-Rabbit IgG, diluted 1/400. Nuclei were counterstained with Hoechst.

    See Abreview

  • ICC/IF image of ab37161 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab37161, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899 Dylight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-HEF1 antibody (ab37161)

This product has been referenced in:
  • Knutson DC  et al. Mice null for NEDD9 (HEF1a) display extensive hippocampal dendritic spine loss and cognitive impairment. Brain Res 1632:141-55 (2016). WB ; Mouse . Read more (PubMed: 26683084) »

See 1 Publication for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (human glioblastoma cell line)
Specification human glioblastoma cell line
Fixative Paraformaldehyde
Permeabilization Yes - 0,1% Triton X-100
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 0.5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Jun 16 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (human glioblastoma cell line)
Specification human glioblastoma cell line
Fixative Paraformaldehyde
Permeabilization Yes - Tritonx100 0.1% in PBS
Blocking step BSA as blocking agent for 20 minute(s) · Concentration: 0.5% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted May 10 2011

Abreviews
Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (neural stem cells)
Loading amount 15 µg
Specification neural stem cells
Gel Running Conditions Reduced Denaturing (7.5%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Ms. Melina Reisenberg

Verified customer

Submitted Jun 23 2009

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Frontal cortex, brain)
Specification Frontal cortex, brain
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Dec 24 2008

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (brain)
Specification brain
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Dec 24 2008

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Tumour: Paget's disease of the nipple)
Specification Tumour: Paget's disease of the nipple
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid pH6
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: rt°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Dec 24 2008

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