Phospho-ErbB2/HER2 (Tyr1248) Antibody [F22D17]

Application: Reactivity: Rat, Human

Usage Information

Dilution
WB1:500 - 1:3000 IHC1:100 - 1:1000

Application
WB, IHC

Reactivity
Rat, Human

Source
Rabbit 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
138 kDa

Positive Control	A431 cells (EGF, 100 ng/mL, 5 min; starved, 16 h); Rat testis; Lung tissue
Negative Control	A431 cells (starved, 16 h)

Datasheet & SDS

Biological Description

Specificity
Phospho-ErbB2/HER2 (Tyr1248) Antibody [F22D17] detects endogenous levels of total ErbB2/HER2 protein only when it is phosphorylated at Tyr1248.

Clone
F22D17

Synonym(s)
CD340; HER2; MLN19; NEU; NGL; ERBB2; Metastatic lymph node gene 19 protein; Proto-oncogene Neu; Proto-oncogene c-ErbB-2; p185erbB2; MLN 19

Background
Phospho-ErbB2/HER2 (Tyr1248) refers to the phosphorylated form of ErbB2 (HER2) at tyrosine 1248, a critical autophosphorylation site within the C-terminal tail of this ligand-independent receptor tyrosine kinase. HER2, notable for its oncogenicity when overexpressed, contains an extracellular ligand-binding domain, a single transmembrane helix, and an intracellular kinase domain with multiple regulatory tyrosines. Phosphorylation at Tyr1248 creates a docking site for adaptor proteins such as SHC and Grb2, activating the Ras-Raf-MAPK pathway to promote proliferation, survival, migration, and angiogenesis, while simultaneously enhancing anti-apoptotic PI3K/AKT signaling. HER2 preferentially heterodimerizes with other ErbB receptors (EGFR/HER3/HER4) or forms homodimers when overexpressed, leading to constitutive signaling. Tyr1248 phosphorylation is strongly associated with HER2 gene amplification and active oncogenic signaling, particularly in aggressive breast, gastric, and ovarian cancers, correlating with poor prognosis and metastatic potential. This site is a target of therapeutic antibodies designed to block HER2-driven transformation.

Background

Phospho-ErbB2/HER2 (Tyr1248) refers to the phosphorylated form of ErbB2 (HER2) at tyrosine 1248, a critical autophosphorylation site within the C-terminal tail of this ligand-independent receptor tyrosine kinase. HER2, notable for its oncogenicity when overexpressed, contains an extracellular ligand-binding domain, a single transmembrane helix, and an intracellular kinase domain with multiple regulatory tyrosines. Phosphorylation at Tyr1248 creates a docking site for adaptor proteins such as SHC and Grb2, activating the Ras-Raf-MAPK pathway to promote proliferation, survival, migration, and angiogenesis, while simultaneously enhancing anti-apoptotic PI3K/AKT signaling. HER2 preferentially heterodimerizes with other ErbB receptors (EGFR/HER3/HER4) or forms homodimers when overexpressed, leading to constitutive signaling. Tyr1248 phosphorylation is strongly associated with HER2 gene amplification and active oncogenic signaling, particularly in aggressive breast, gastric, and ovarian cancers, correlating with poor prognosis and metastatic potential. This site is a target of therapeutic antibodies designed to block HER2-driven transformation.

References
https://pubmed.ncbi.nlm.nih.gov/21712485/ https://www.explorationpub.com/Journals/em/Article/1001237

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%