HEF1/NEDD9 Antibody [M16B3] | Primary Antibodies

Application: Reactivity: Human, Mouse, Rat, Monkey

Lane 1: A549, Lane 2: C2C12

Usage Information

Dilution
WB1:1000 IP1:100

Application
WB, IP

Reactivity
Human, Mouse, Rat, Monkey

Source
Mouse Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
105 kDa, 115 kDa

Positive Control	A549 cells; C2C12 cells
Negative Control

Datasheet & SDS

Biological Description

Specificity
HEF1/NEDD9 Antibody [M16B3] detects endogenous levels of total HEF1/NEDD9 protein.

Clone
M16B3

Synonym(s)
NEDD9; hEF1; CRK-associated substrate-related protein (CAS-L; CasL); Cas scaffolding protein family member 2 (CASS2); NEDD-9; Renal carcinoma antigen NY-REN-12; p105; Enhancer of filamentation 1 p55; CASL

Background
HEF1, also known as NEDD9 or Cas-L, is a key multidomain adaptor protein within the Cas family, coordinating integrin-mediated signaling at focal adhesions during interphase and localizing to centrosomes and mitotic structures in G2/M phase to regulate cell adhesion, migration, polarity, and survival. HEF1 contains an N-terminal SH3 domain (residues 3–65) for binding proline-rich proteins, a substrate domain (residues 110–356) enriched in tyrosine motifs such as Y189 and Y214 that are phosphorylated by Src and FAK to recruit SH2 domain-containing effectors like Crk and Abl, a serine-rich region (residues 357–560) forming a four-helix bundle for structural stability and protein interactions, and a C-terminal focal adhesion targeting (FAT) domain (residues 561–834) with helix-loop-helix motifs that facilitate FAK docking, Src binding, and dimerization. HEF1 integrates signals from FAK, Src, PDGFR, and Abl, leading to phosphorylation of its substrate domain and assembly of signaling complexes that promote actin cytoskeleton remodeling, focal adhesion turnover, invadopodia formation via MMP9 secretion, and activation of pathways such as FAK-Src-Aurora-A for chemotaxis and mitotic progression; its phosphorylation dynamics further regulate apoptosis resistance and epithelial-mesenchymal transition. NEDD9 overexpression drives metastasis in cancers like glioblastoma, melanoma, and lung carcinoma by enhancing invasion and survival signaling, while deficiency impairs tumor progression and leukocyte migration.

Background

HEF1, also known as NEDD9 or Cas-L, is a key multidomain adaptor protein within the Cas family, coordinating integrin-mediated signaling at focal adhesions during interphase and localizing to centrosomes and mitotic structures in G2/M phase to regulate cell adhesion, migration, polarity, and survival. HEF1 contains an N-terminal SH3 domain (residues 3–65) for binding proline-rich proteins, a substrate domain (residues 110–356) enriched in tyrosine motifs such as Y189 and Y214 that are phosphorylated by Src and FAK to recruit SH2 domain-containing effectors like Crk and Abl, a serine-rich region (residues 357–560) forming a four-helix bundle for structural stability and protein interactions, and a C-terminal focal adhesion targeting (FAT) domain (residues 561–834) with helix-loop-helix motifs that facilitate FAK docking, Src binding, and dimerization. HEF1 integrates signals from FAK, Src, PDGFR, and Abl, leading to phosphorylation of its substrate domain and assembly of signaling complexes that promote actin cytoskeleton remodeling, focal adhesion turnover, invadopodia formation via MMP9 secretion, and activation of pathways such as FAK-Src-Aurora-A for chemotaxis and mitotic progression; its phosphorylation dynamics further regulate apoptosis resistance and epithelial-mesenchymal transition. NEDD9 overexpression drives metastasis in cancers like glioblastoma, melanoma, and lung carcinoma by enhancing invasion and survival signaling, while deficiency impairs tumor progression and leukocyte migration.

References
https://pubmed.ncbi.nlm.nih.gov/17703068/ https://pubmed.ncbi.nlm.nih.gov/17908996/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%