Phospho-Stat5 (Tyr694) Antibody [L6J15]

Application: Reactivity: Human, Mouse

Usage Information

Dilution
WB1:1000 IHC1:200 - 1:800 IF1:100 - 1:400 FCM1:100 - 1:400

Application
WB, IHC, IF, FCM

Reactivity
Human, Mouse

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
90 kDa

Positive Control	Human breast carcinoma; Human lung carcinoma; TF-1 cells( hGM-CSF, 50 ng/mL, 15 min); HeLa cells (IFNα-treated); K562 (Gleevec, 1 h)
Negative Control	TF-1 cells; HeLa cells; K562 (STI-571–treated)

Datasheet & SDS

Biological Description

Specificity
Phospho-Stat5 (Tyr694) Antibody [L6J15] detects endogenous levels of total Stat5 protein only when it is phosphorylated at Tyr694.

Clone
L6J15

Synonym(s)
Signal transducer and activator of transcription 5A; STAT5A; STAT5; Signal transducer and activator of transcription 5B; STAT5B

Background
Phospho-Stat5 (Tyr694) represents the activated form of Signal Transducer and Activator of Transcription 5 (STAT5), a member of the STAT family of cytoplasmic transcription factors involved in mediating cytokine signaling. STAT5 has two major isoforms: STAT5A, which is phosphorylated at Tyr694, and STAT5B, phosphorylated at Tyr699. Phosphorylation at Tyr694 is essential for STAT5 activation and is mediated by kinases such as JAK2, Src family kinases, and others in response to a diverse array of ligands, including IL-2, IL-3, IL-7, GM-CSF, erythropoietin, thrombopoietin, growth hormones, stem cell factor (KITLG/SCF), and ERBB4. This phosphorylation event enables STAT5 dimerization, nuclear translocation, and binding to gamma-activated sequence (GAS) elements in DNA, thereby driving transcriptional activation. Activated phospho-STAT5 is crucial for signal transduction and gene regulation, controlling cellular proliferation, differentiation, survival, lactation (such as PRL-induced milk protein gene expression), immune responses, hematopoiesis, angiogenesis, cell motility, and somatic growth. It operates in key cytokine-mediated pathways, including the JAK-STAT cascade, growth factor signaling via FGFR1-4, and interferon responses, with STAT5A and STAT5B isoforms regulated independently across different cell types, notably endothelial and hematopoietic cells. In disease, constitutive or stimulus-independent phospho-STAT5 activation, often via TEL/JAK2 gene fusions, is a driver of tumorigenesis in malignancies such as myeloma and lymphoma.

Background

Phospho-Stat5 (Tyr694) represents the activated form of Signal Transducer and Activator of Transcription 5 (STAT5), a member of the STAT family of cytoplasmic transcription factors involved in mediating cytokine signaling. STAT5 has two major isoforms: STAT5A, which is phosphorylated at Tyr694, and STAT5B, phosphorylated at Tyr699. Phosphorylation at Tyr694 is essential for STAT5 activation and is mediated by kinases such as JAK2, Src family kinases, and others in response to a diverse array of ligands, including IL-2, IL-3, IL-7, GM-CSF, erythropoietin, thrombopoietin, growth hormones, stem cell factor (KITLG/SCF), and ERBB4. This phosphorylation event enables STAT5 dimerization, nuclear translocation, and binding to gamma-activated sequence (GAS) elements in DNA, thereby driving transcriptional activation. Activated phospho-STAT5 is crucial for signal transduction and gene regulation, controlling cellular proliferation, differentiation, survival, lactation (such as PRL-induced milk protein gene expression), immune responses, hematopoiesis, angiogenesis, cell motility, and somatic growth. It operates in key cytokine-mediated pathways, including the JAK-STAT cascade, growth factor signaling via FGFR1-4, and interferon responses, with STAT5A and STAT5B isoforms regulated independently across different cell types, notably endothelial and hematopoietic cells. In disease, constitutive or stimulus-independent phospho-STAT5 activation, often via TEL/JAK2 gene fusions, is a driver of tumorigenesis in malignancies such as myeloma and lymphoma.

References
https://pubmed.ncbi.nlm.nih.gov/11641791/ https://pubmed.ncbi.nlm.nih.gov/39191751/

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%