NEDD9/CASL/HEF1 Antibody (2G9)

Images

 
Western Blot: NEDD9/CASL/HEF1 Antibody (2G9) [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 ...read more
Immunocytochemistry/ Immunofluorescence: NEDD9/CASL/HEF1 Antibody (2G9) [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) [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 ...read more

Product Details

Summary
Reactivity Hu, MuSpecies Glossary
Applications WB, ELISA, ICC/IF, IP
Clone
2G9
Clonality
Monoclonal
Host
Mouse
Conjugate
Unconjugated
Concentration
1 mg/ml

NEDD9/CASL/HEF1 Antibody (2G9) Summary

Immunogen
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).
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
Does not recognize p130 Cas. Not tested on Sin1.This mostly detects HEF1 localized at the focal adhesion sites.
Isotype
IgG1 Kappa
Clonality
Monoclonal
Host
Mouse
Gene
NEDD9
Purity
Protein A purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:5000
  • ELISA 1:5000-1:20000
  • Immunocytochemistry/Immunofluorescence 1:500
  • Immunoprecipitation 1:1000
Application Notes
This monoclonal antibody has been tested for use in western blotting, immunoprecipitation and Immunofluorescence. Expect bands approximately 115 and 105 in size corresponding - 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.
Publications
Read Publications using
NB100-1699 in the following applications:

Reactivity Notes

Cross reactivity with NEDD9/CASL/HEF1 from other sources has not been tested.

Packaging, Storage & Formulations

Storage
Store at -20C. Avoid freeze-thaw cycles.
Buffer
0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2
Preservative
0.01% Sodium Azide
Concentration
1 mg/ml
Purity
Protein A purified

Alternate Names for NEDD9/CASL/HEF1 Antibody (2G9)

  • Cas scaffolding protein family member 2
  • CAS2
  • CASL
  • CAS-LCASS2cas-like docking
  • CASLdJ49G10.2
  • CRK-associated substrate-related protein
  • dJ49G10.2 (Enhancer of Filamentation 1 (HEF1))
  • dJ761I2.1 (enhancer of filamentation (HEF1))
  • dJ761I2.1
  • enhancer of filamentation 1
  • HEF1
  • HEF1Crk-associated substrate related
  • NEDD9
  • NEDD-9
  • Neural precursor cell expressed developmentally down-regulated protein 9
  • neural precursor cell expressed, developmentally down-regulated 9
  • p105
  • Renal carcinoma antigen NY-REN-12

Background

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.

Limitations

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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Applications: WB, ELISA, ICC/IF, IP

Publications for NEDD9/CASL/HEF1 Antibody (NB100-1699)(3)

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ICC/IF Video Protocol

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Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 350 NB100-1699AF350
Alexa Fluor 405 NB100-1699AF405
Alexa Fluor 488 NB100-1699AF488
Alexa Fluor 532 NB100-1699AF532
Alexa Fluor 594 NB100-1699AF594
Alexa Fluor 647 NB100-1699AF647
Alexa Fluor 700 NB100-1699AF700
Alexa Fluor 750 NB100-1699AF750
DyLight 350 NB100-1699UV
DyLight 405 NB100-1699V
DyLight 488 NB100-1699G
DyLight 550 NB100-1699R
DyLight 594 NB100-1699DL594
DyLight 680 NB100-1699FR
DyLight 755 NB100-1699IR
Janelia Fluor 549 NB100-1699JF549
Janelia Fluor 646 NB100-1699JF646

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Bioinformatics

Gene Symbol NEDD9
Entrez
Uniprot