STAT5a Recombinant Rabbit Monoclonal Antibody [SC66-09]
Overview
Product Name
STAT5a Recombinant Rabbit Monoclonal Antibody [SC66-09]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within C-terminal human STAT5a.
Product Specificity
Stat5a [SC66-09] Recombinant Rabbit Antibody recognizes endogenous levels of total Stat5a protein. This antibody does not cross-react with Stat5b protein.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Molecular Weight
Predicted band size: 91 kDa
Positive Control
HeLa cell lysate, K-562 cell lysate, RAW264.7 cell lysate, EL4 cell lysate, Rat spleen tissue lysate, mouse bone tissue, mouse spleen tissue, rat bone tissue, rat spleen tissue, human tonsil tissue, Hela, C2C12, RAW264.7.
Conjugation
unconjugated
Clone Number
SC66-09
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:1,000-1:2,000
-
IF-Cell
-
1:50-1:200
-
IF-Tissue
-
1:50-1:200
-
IHC-P
-
1:200-1:1,000
-
IP
-
1-2μg/sample
-
FC
-
1:1,000
Target
Function
Signal transducer and activator of transcription 5A (Stat5a) and Stat5b, which share 96% homology, undergo receptor tyrosine kinase or G protein-coupled receptor-dependent phosphorylation in response to cytokines or growth factors, and then form homo- or heterodimers that translocate to the nucleus, where they initiate transcription. Activation of Stat5a via IL-2, IL-3, IL-7/ GM-CSF, erythropoietin, thrombopoietin and growth hormones influences proliferation, differentiation and apoptosis in lymphohematopoietic cells. Phos-phorylation of Stat5a at Ser127/Ser128 and Ser779 are contigent on ErbB-4-mediated activation of Stat5a. Activation of Stat5b via IL-2, IL-4, CSF-1 and growth hormones influences TCR signaling, apoptosis, adult mammary gland development and sexual dimorphism of liver gene expression. Stat5b is the major liver-expressed Stat5 form that has been shown to fuse with the retin-oic acid receptor a gene in acute promyelocytic leukemias (APLL). Stat5a/b null mice have severely impaired lymphoid development and differentiation.
Background References
1. Cho KI et al. Differential loss of prolyl isomerase or chaperone activity of Ran-binding protein 2 (Ranbp2) unveils distinct physiological roles of its cyclophilin domain in proteostasis. J Biol Chem 289:4600-25 (2014).
2. Surana R et al. IL4 limits the efficacy of tumor-targeted antibody therapy in a murine model. Cancer Immunol Res 2:1103-12 (2014).
Sequence Similarity
Belongs to the transcription factor STAT family.
Post-translational Modification
Tyrosine phosphorylated in response to KITLG/SCF, IL2, IL3, IL7, IL15, CSF2/GMCSF, GH1, PRL, EPO and THPO (By similarity). Activated KIT promotes phosphorylation on tyrosine residues and subsequent translocation to the nucleus. Tyrosine phosphorylated in response to constitutively activated FGFR1, FGFR2, FGFR3 and FGFR4 (By similarity). Tyrosine phosphorylation is required for DNA-binding activity and dimerization. Serine phosphorylation is also required for maximal transcriptional activity (By similarity). Tyrosine phosphorylated in response to signaling via activated FLT3; wild-type FLT3 results in much weaker phosphorylation than constitutively activated mutant FLT3. Alternatively, can be phosphorylated by JAK2 at Tyr-694.; ISGylated.
Subcellular Location
Nucleus, Cytoplasm.
Synonyms
Mammary gland factor antibody
MGF antibody
Signal transducer and activator of transcription 5A antibody
STA5A_HUMAN antibody
STAT 5 antibody
STAT 5A antibody
STAT5 antibody
STAT5A antibody
Images
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Western blot analysis of STAT5a on different lysates with Rabbit anti-STAT5a antibody (ET1610-58) at 1/1,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: K-562 cell lysate
Lane 3: RAW264.7 cell lysate
Lane 4: EL4 cell lysate
Lane 5: Rat spleen tissue lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 91 kDa
Observed band size: 91 kDa
Exposure time: 10 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1610-58) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
All lanes: Western blot analysis of STAT5a with anti-STAT5a antibody (ET1610-58) at 1/500 dilution.
Lane 1: Wild-type Hela whole cell lysate.
Lane 2/3: STAT5a knockdown Hela whole cell lysate.
ET1610-58 was shown to specifically react with STAT5a in Wild-type Hela cells. Weakened bands were observed when STAT5a knockdown samples were tested. Wild-type and STAT5a knockdown samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary Anti-STAT5a antibody (ET1610-58, 1/500) and Anti-HSP90 antibody (ET1605-56, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG H&L (HRP) Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded mouse bone tissue with Rabbit anti-STAT5a antibody (ET1610-58) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-58) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-STAT5a antibody (ET1610-58) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-58) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat bone tissue with Rabbit anti-STAT5a antibody (ET1610-58) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-58) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-STAT5a antibody (ET1610-58) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-58) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-STAT5a antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1610-58, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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ICC staining of STAT5a in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-58, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of STAT5a in C2C12 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1610-58, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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Flow cytometric analysis of RAW264.7 cells labeling STAT5a.
Cells were fixed and permeabilized. Then stained with the primary antibody (ET1610-58, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). -
STAT5a was immunoprecipitated from 0.2 mg RAW264.7 cell lysate with ET1610-58 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1610-58 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: RAW264.7 cell lysate (input)
Lane 2: ET1610-58 IP in RAW264.7 cell lysate
Lane 3: Rabbit IgG instead of ET1610-58 in RAW264.7 cell lysate
Blocking/Dilution buffer: primary antibody dilution (K1803)
Exposure time: 46 seconds; ECL: K1801
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Citation
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Exercise ameliorates hepatic lipid accumulation via upregulating serum PRL and activating hepatic PRLR-mediated JAK2/STAT5 signaling pathway in NAFLD mice
Journal: Frontiers In Pharmacology
DOI: 10.3389/fphar.2025.1647231
IF: 4.8
Application: WB
Reactivity: Mouse
Publish date: 2025 Aug
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Intracellular angiopoietin-1 promotes TKI-resistance via activation of JAK/STAT5 pathway in chronic myeloid leukemia
Journal: Oncogene
DOI:
IF: 8
Application: WB
Reactivity: Human
Publish date: 2023 Jan
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Therapeutic potential of icariin in rats with letrozole and high-fat diet-induced polycystic ovary syndrome.
Journal:
DOI:
IF: 5.195
Application: WB
Reactivity: Rat
Publish date: 2023 Aug
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