
Recombinant Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
- Datasheet
- References (2)
- Protocols
Overview
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Product name
Anti-STAT5a + STAT5b antibody [EPR16671-40] -
Description
Rabbit monoclonal [EPR16671-40] to STAT5a + STAT5b -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, ICC/IF, Flow Cyt, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment within Mouse STAT5a aa 600 to the C-terminus. The exact sequence is proprietary. Also SwissProt ID: P42232 (mouse)
Database link: P42230 -
Positive control
- WB: Human STAT5A full length recombinant protein; K562, Jurkat and Daudi cell lysates; Human fetal heart and kidney lysates; mouse heart, kidney, spleen lysates; rat brain, heart, spleen lysates; RAW 264.7, PC12, NIH/3T3 whole cell lysates. IHC-P: Human tonsils, mouse spleen and rat spleen tissue. IF, Flow, IP: K562 cells
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General notes
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
This product is a recombinant rabbit monoclonal antibody.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR16671-40 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab194898 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IHC-P | 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. | |
WB | 1/2000. Detects a band of approximately 91 kDa (predicted molecular weight: 91 kDa). | |
ICC/IF | 1/250. | |
Flow Cyt | 1/80. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
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IP | 1/100. |
Target
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Cellular localization
STAT5a: Cytoplasm. Nucleus. Translocated into the nucleus in response to phosphorylation. STAT5b: Cytoplasm. Nucleus. Translocated into the nucleus in response to phosphorylation. -
Database links
- Entrez Gene: 6776 Human
- Entrez Gene: 6777 Human
- Entrez Gene: 20850 Mouse
- Entrez Gene: 20851 Mouse
- Entrez Gene: 24918 Rat
- Entrez Gene: 25126 Rat
- Omim: 601511 Human
- Omim: 604260 Human
see all
Images
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All lanes : Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898) at 1/20000 dilution
Lane 1 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysates
Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates
Lane 3 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 91 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling STAT5a + STAT5b with ab194898 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/400 dilution (green). Confocal image showing both cytoplasmic and nuclear staining on K562 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor® 594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
1. ab194898 at 1/250 dilution followed by ab150120 (Alexa Fluor® 594 Goat anti-Mouse secondary) at 1/500 dilution.
2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor® 488 Goat Anti-Rabbit IgG H&L) at 1/400 dilution. -
All lanes : Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898) at 1/2000 dilution
Lane 1 : Human fetal heart lysates
Lane 2 : Human fetal kidney lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 91 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898) at 1/1000 dilution + Human STAT5A full length recombinant protein at 10 µg
Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 91 kDaRecombinant human C-terminal Myc/DDK tagged full length STAT5A from a commercial source.
Blocking/Dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898) at 1/2000 dilution
Lane 1 : mouse heart lysates
Lane 2 : mouse kidney lysates
Lane 3 : mouse spleen lysates
Lane 4 : rat brain lysates
Lane 5 : rat heart lysates
Lane 6 : rat spleen lysates
Lane 7 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysates
Lane 8 : PC12 (Rat adrenal gland pheochromocytoma) whole cell lysates
Lane 9 : NIH3T3 (Mouse embyro fibroblast cells) whole cell lysates
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 91 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling STAT5a + STAT5b using ab194898 at 1/2000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 was used as secondary. Counter stained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab194898.
Nuclear and weak cytoplasmic staining on lymphocytes of human tonsil is observed.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling STAT5a + STAT5b using ab194898 at 1/2000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 was used as secondary. Counter stained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab194898.
Nuclear and weak cytoplasmic staining on lymphocytes of mouse spleen is observed.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)
Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling STAT5a + STAT5b using ab194898 at 1/2000 dilution. A Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 was used as secondary. Counter stained with Hematoxylin.
Inset image: negative control obtained using PBS instead of ab194898.
Nuclear and weak cytoplasmic staining on lymphocytes of rat spleen is observed.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Flow cytometric analysis of 2% paraformaldehyde-fixed K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling STAT5a + STAT5b with ab194898 at 1/80 dilution (red) compared with a rabbit monoclonal IgG isotype control (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
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STAT5a and STAT5b were immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) cells with ab194898 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab194898 at 1/1000 dilution. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated, was used as secondary antibody at 1/1000 dilution.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
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STAT5a + STAT5b were co-immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) cells with ab194898 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab32043 (Rabbit monoclonal [E289] to STAT5a) at 1/1000 dilution. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated, was used as secondary antibody at 1/1000 dilution.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
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STAT5a + STAT5b were co-immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) cells with ab194898 at 1/100 dilution. Western blot was performed from the immunoprecipitate using ab178941 (Rabbit monoclonal [EPR16671] to STAT5b) at 1/1000 dilution. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated, was used as secondary antibody at 1/1000 dilution.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Protocols
References
This product has been referenced in:
- Ding L et al. Deletion of Cdkn1b in ACI rats leads to increased proliferation and pregnancy-associated changes in the mammary gland due to perturbed systemic endocrine environment. PLoS Genet 15:e1008002 (2019). Read more (PubMed: 30893315) »
- Mitchell J et al. Validation of a Preclinical Model of Diethylnitrosamine-Induced Hepatic Neoplasia in Yucatan Miniature Pigs. Oncology 91:90-100 (2016). Read more (PubMed: 27305144) »