Recombinant Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

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Western blot - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR16668] to STAT5a + STAT5b
Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WB
Reacts with: Mouse, Rat, Human, Recombinant fragment

Product name

Anti-STAT5a + STAT5b antibody [EPR16668]
See all STAT5a+STAT5b primary antibodies
Description

Rabbit monoclonal [EPR16668] to STAT5a + STAT5b
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), ICC/IF, IHC-P, WBmore details
Species reactivity

Reacts with: Mouse, Rat, Human, Recombinant fragment
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Full length human Stat5a recombinant protein; recombinant protein fragment mouse stat5b; K562, Jurkat, Daudi, C6, PC-12 and NIH/3T3 whole cell lysates; mouse and rat heart lysates. IHC-P: Human tonsil, mouse liver and rat cardiac muscle tissues. ICC/IF: RAW 264.7 and C6 cells. Flow Cyt (intra): NIH/3T3 cells.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR16668
Isotype

IgG
Research areas

Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (ab210849)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab200341 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
Flow Cyt (Intra)		1/80.
ICC/IF		1/500.
IHC-P		1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB		1/200 - 1/10000. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa).

Notes
Flow Cyt (Intra) 1/80.
ICC/IF 1/500.
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/200 - 1/10000. Detects a band of approximately 90 kDa (predicted molecular weight: 90 kDa).

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 [EPR16668] (ab200341)

Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341) at 1/200 dilution + Full length human Stat5a recombinant protein at 0.01 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 90 kDa
Observed band size: 90 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
ab200341 show cross reactivity with STAT5a. Full length human Stat5a recombinant protein contains aa1-794 with MYC/DDDDK tag.
Western blot - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341) at 1/10000 dilution + Recombinant protein fragment mouse Stat5b at 0.01 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 90 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

All lanes : Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341) at 1/1000 dilution

Lane 1 : K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
Lane 3 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysate

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: 90 kDa
Observed band size: 90 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

All lanes : Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341) at 1/1000 dilution

Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
Lane 2 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 3 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate
Lane 4 : Mouse heart lysate
Lane 5 : Rat heart lysate

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: 90 kDa
Observed band size: 90 kDa

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling STAT5b with ab200341 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

Cytoplasm and nuclear staining on Human tonsil tissue is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling STAT5b with ab200341 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

Cytoplasm and nuclear staining on mouse liver tissue is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

Immunohistochemical analysis of paraffin-embedded Rat cardiac muscle tissue labeling STAT5b with ab200341 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

Cytoplasm and nuclear staining on rat cardiac muscle tissue is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) cells labeling STAT5b with ab200341 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).
Nuclear and cytoplasm staining on RAW 264.7 cell line is observed.
The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab200341 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (Rat glial tumor cells) cells labeling STAT5b with ab200341 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).
Nuclear and cytoplasm staining on C6 cell line is observed.
The nuclear counter stain is DAPI (blue).
Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab200341 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.
Flow Cytometry (Intracellular) - Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed NIH/3T3 (Mouse embyro fibroblast cells) cells labeling STAT5b with ab200341 at 1/80 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730;black) and an 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.
Anti-STAT5a + STAT5b antibody [EPR16668] (ab200341)

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

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab200341? Please let us know so that we can cite the reference in this datasheet.

ab200341 has been referenced in 1 publication.

Wang S et al. Long non-coding RNA SNHG1 suppresses cell migration and invasion and upregulates SOCS2 in human gastric carcinoma. Biochem Biophys Rep 27:101052 (2021). PubMed: 34179518

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Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Compatible Secondaries

Isotype control

Applications

The Abpromise guarantee

Target

Cellular localization

Database links

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (1)

Customer reviews and Q&As