製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to HEF1
  • アプリケーション
    適用あり: WB, ICC/IFmore details
  • 種交差性
    交差種: Human
    交差が予測される動物種: Mouse, Rat
  • 免疫原

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

    (Peptide available as ab39117.)

  • ポジティブ・コントロール
    • 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)

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
WB 1/250. Detects a band of approximately 120 kDa (predicted molecular weight: 110 kDa).
ICC/IF Use a concentration of 5 µg/ml.

ターゲット情報

  • 機能
    Docking 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.
  • 組織特異性
    Widely 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.
  • 配列類似性
    Belongs to the CAS family.
    Contains 1 SH3 domain.
  • ドメイン
    Contains 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.
  • 翻訳後修飾
    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.
  • 細胞内局在
    Cytoplasm > 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
  • 参照データベース
  • 別名
    • 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

画像

  • 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.

参考文献

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) »
  • Ahn J  et al. The metastasis gene NEDD9 product acts through integrin ß3 and Src to promote mesenchymal motility and inhibit amoeboid motility. J Cell Sci 125:1814-26 (2012). Read more (PubMed: 22328516) »

See all 2 Publications for this product

レビューと Q&A

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

投稿 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

投稿 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

投稿 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

投稿 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

投稿 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

投稿 Dec 24 2008

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

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