Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Mouse

Applications

Validated:

ELISA, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Paraffin, Immunoprecipitation, Western Blot

Cited:

Immunocytochemistry/ Immunofluorescence, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 2G9

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

View all NEDD9/CASL Antibodies »

Product Specifications

Immunogen

NEDD9/CASL/HEF1 Antibody (2G9) was produced by repeated immunizations with a synthetic peptide corresponding to amino acid residues 82-398 of human NEDD9/CASL/HEF1 protein (hNEDD9/CASL/HEF1, 843 aa, predicted MW 92.8 kDa). (Uniprot: Q14511)

Reactivity Notes

Reactivity against homologues from other sources is not known

Localization

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 predomin

Specificity

This Protein A purified antibody is directed against human HEF1 protein. Reactivity occurs against human, mouse and rat forms of the protein. Reactivity against multiple isoforms is expected. Reactivity against homologues from other sources is not known. Specificity was determined by partial epitope mapping.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Description

This Protein A purified antibody is directed against human NEDD9/CASL/HEF1 protein. The product was purified from tissue culture supernatant by chromatography

Store vial at -20C prior to opening. Aliquot contents and freeze at -20C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4C as an undiluted liquid. Dilute only prior to immediate use.

Scientific Data Images for NEDD9/CASL/HEF1 Antibody (2G9) - (82-398 aa)

Western Blot: NEDD9/CASL/HEF1 Antibody (2G9)(82-398 aa) [NB100-1699]

Western Blot: NEDD9/CASL/HEF1 Antibody (2G9)(82-398 aa) [NB100-1699]

Western Blot: NEDD9/CASL/HEF1 Antibody (2G9) - (82-398 aa) [NB100-1699] - Detection of a 115 kDa band corresponding to HEF1 in MCF7 lysate (arrowhead). Approximately 35 ig 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 IRDye800 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). IRDye800 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.
Immunocytochemistry/ Immunofluorescence: NEDD9/CASL/HEF1 Antibody (2G9) - (82-398 aa) [NB100-1699]

Immunocytochemistry/ Immunofluorescence: NEDD9/CASL/HEF1 Antibody (2G9) - (82-398 aa) [NB100-1699]

Immunocytochemistry/Immunofluorescence: NEDD9/CASL/HEF1 Antibody (2G9) - (82-398 aa) [NB100-1699] - Detection of HEF1 localized at focal adhesion sites. The antibody was used at a 1:500 dilution with a 3-sec exposure time.
Western Blot: NEDD9/CASL/HEF1 Antibody (2G9)(82-398 aa) [NB100-1699]

Western Blot: NEDD9/CASL/HEF1 Antibody (2G9)(82-398 aa) [NB100-1699]

Western Blot: NEDD9/CASL/HEF1 Antibody (2G9) - (82-398 aa) [NB100-1699] - Detection of endogenous HEF1 present in various cell lines. Panel A shows detection using a 15 min exposure. Panel B is the same blot exposed for 2 min. The doublet represents p105 and p115 staining. The lower MW band represents p55. 3Y1 cells are derived from rat fibroblast cells. Mouse 3T3 cells are also reactive (not shown). To date no staining has been noted on CHO cells.
View 1 more images

Applications for NEDD9/CASL/HEF1 Antibody (2G9) - (82-398 aa)

Application
Recommended Usage

ELISA

1:5000-1:20000

Immunocytochemistry/ Immunofluorescence

1:500

Immunohistochemistry

1:10-1:500

Immunohistochemistry-Paraffin

1:200-1:500

Immunoprecipitation

1:1000

Western Blot

1:5000
Application Notes
This monoclonal antibody has been tested for use in western blotting, immunoprecipitation and immunofluorescence. Specific conditions for reactivity should be optimized by the end user. Expect bands approximately 115 and 105 in size corresponding to isoforms of HEF1 protein by western blotting in the appropriate cell lysate or extract. This antibody does not recognize p130Cas. Sin1 has not been tested. IF was performed using 4% PFA fixed cells. This monoclonal antibody mostly detects HEF1 localized at the focal adhesion sites.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2

Preservative

0.01% Sodium Azide

Concentration

Please see the vial label for concentration. If unlisted please contact technical services.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at -20C short term. Aliquot and store at -80C long term. Avoid freeze-thaw cycles.

Background: NEDD9/CASL

HEF1, also known as Enhancer of filamentation 1, CRKassociated substrate-related protein, CAS-L, CasL, p105 and Neural precursor cell expressed developmentally down-regulated 9 is the product of the NEDD9 (CASGL) gene. HEF1 functions as a docking protein that plays a central coordinating role for tyrosine-kinase-based signaling related to cell adhesion. HEF1 may also 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. HEF1 may also 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. HEF1 forms a homodimer and can heterodimerize with HLH proteins ID2, E12, E47 and also with p130cas. HEF1 also forms complexes in vivo with related adhesion focal tyrosine kinase (RAFTK), adapter protein CRKL and LYN kinase and also interacts with MICAL and TXNL4/DIM1. This protein localizes to both the cell nucleus and the cell periphery and is differently localized in fibroblasts and epithelial cells. In fibroblasts, it is predominantly nuclear and in some cells is present in the Golgi apparatus. In epithelial cells, it is localized predominantly in the cell periphery with particular concentration in lamellipodia, but it is also found in the nucleus. HEF1 is widely expressed although higher levels are detected in kidney, lung, and placental tissue. HEF1 is also detected in T-cells, B-cells and diverse cell lines. HEF1 is activated upon induction of cell growth. 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. Isoforms p105 and p115 are predominantly cytoplasmic and associate with focal adhesions while p55 associates with the mitotic spindle.

Long Name

Neural Precursor Cell Expressed Developmentally Down-Regulated Protein 9

Alternate Names

CAS2, CASL, HEF1, p105

Entrez Gene IDs

4739 (Human); 18003 (Mouse); 291044 (Rat)

Gene Symbol

NEDD9

UniProt

Q14511 (Human)

Additional NEDD9/CASL Products

Product Documents for NEDD9/CASL/HEF1 Antibody (2G9) - (82-398 aa)

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

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Product Specific Notices for NEDD9/CASL/HEF1 Antibody (2G9) - (82-398 aa)

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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