Recombinant Anti-STAT5a + STAT5b antibody [EPR16671-40] - BSA and Azide free (ab215367)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR16671-40] to STAT5a + STAT5b - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-STAT5a + STAT5b antibody [EPR16671-40] - BSA and Azide free -
Description
Rabbit monoclonal [EPR16671-40] to STAT5a + STAT5b - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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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
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General notes
ab215367 is the carrier-free version of ab194898.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
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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Conjugation kits
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab215367 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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
Use at an assay dependent concentration. Detects a band of approximately 91 kDa (predicted molecular weight: 91 kDa).
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IHC-P | (1) |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Detects a band of approximately 91 kDa (predicted molecular weight: 91 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
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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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab194898).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab194898).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab194898).
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab194898).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab194898).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab194898).
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Intracellular 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab194898).
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This IHC data was generated using the same anti-STAT5a/b antibody clone, EPR16671-40, in a different buffer formulation (cat# 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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This ICC/IF data was generated using the same anti-STAT5a/b antibody clone, EPR16671-40, in a different buffer formulation (cat# 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.
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (1)
ab215367 has been referenced in 1 publication.
- Bi CS et al. Calcitriol inhibits osteoclastogenesis in an inflammatory environment by changing the proportion and function of T helper cell subsets (Th2/Th17). Cell Prolif 53:e12827 (2020). PubMed: 32406154