Anti-STAT5a + STAT5b antibody [EPR16671-40] Recombinant (ab194898)

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

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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
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.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR16671-40
Isotype

IgG
Research areas
- Cell Biology
- Apoptosis
- Nucleus
- Other
- Cancer
- Cell Death
- Apoptosis
- Nucleus
- Other

Alternative Versions
- Anti-STAT5a + STAT5b antibody [EPR16671-40] - BSA and Azide free (ab215367)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

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
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.
IP		1/100.

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
- SwissProt: P42229 Human
- SwissProt: P51692 Human
- SwissProt: P42230 Mouse
- SwissProt: P42232 Mouse
- SwissProt: P52632 Rat
- SwissProt: Q62771 Rat
- Unigene: 437058 Human
- Unigene: 595276 Human
- Unigene: 277403 Mouse
- Unigene: 34064 Mouse
- Unigene: 154399 Rat
- Unigene: 54486 Rat
see all

Western blot - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

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 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunocytochemistry/ Immunofluorescence - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

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.
Western blot - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

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 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

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 kDa

Recombinant human C-terminal Myc/DDK tagged full length STAT5A from a commercial source.
Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

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 kDa

Blocking/Dilution buffer: 5% NFDM/TBST.
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.
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.
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.
Flow Cytometry - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

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.
Immunoprecipitation - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

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.
Immunoprecipitation - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

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.
Immunoprecipitation - Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

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.

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

This product has been referenced in:

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) »

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Application

Western blot

Sample

Human Tissue lysate - whole (human sceletal muscle)

Gel Running Conditions

Reduced Denaturing (WES simple western)

Loading amount

1 µg

Specification

human sceletal muscle

Blocking step

antibody dilution from protein simple as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 23°C

Helle Zibrandtsen

Verified customer

Submitted Mar 07 2018

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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Anti-STAT5a + STAT5b antibody [EPR16671-40] (ab194898)

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Western blot abreview for Anti-STAT5a + STAT5b antibody [EPR16671-40]