STAT5 Total PhosphoTracer ELISA Kit (ab119653)
- Product nameSTAT5 Total PhosphoTracer ELISA Kit
- Detection methodFluorescent
- Tests1 x 96 well plate
- Sample typeCell culture extracts
- Assay typeSemi-quantitative
- Sensitivity10 µg protein/ml
- Range10 µg protein/ml - 1000 µg protein/ml
- Assay time2h 0m
- Assay durationOne step assay
- Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat
- Product overview
PhosphoTracer assays use a traditional immuno-sandwich format, but with a major difference both the analyte and the assay reagents are added to the PhosphoTracer assay microplate at the same time. After a short incubation period, unbound assay reagents and analytes are washed away, and immuno-complexes containing both antibodies are detected. The process can take as little as 60 minutes to complete.
PhosphoTracer kits also allow a higher degree of assay flexibility. In contrast to other ELISA formats, no antibodies are present on the assay microplate itself, so assays for several different targets can be performed in different wells on the same microplate. Simply mix the lysate with your target reagents of choice, using the microplate configuration of your choice.
A whole new way of performing cellular assays, PhosphoTracer takes the hard work out of running a standard ELISA, while still giving the high quality results expected from a sandwich immunoassay. Fully self-contained kits are supplied in convenient 96-well packs. Simple to use and highly sensitive PhosphoTracer kits are designed to get results, fast.
Abcam’s PhosphoTracer phospho-STAT5 assay kits detect endogenous levels of STAT5A and STAT5B (GenBank Accessions NP_003143 and NP_036580) in cellular lysates. Total STAT5 assay kits detect STAT5 irrespective of phosphorylation.
The substrate used with the HRP conjugated detection antibody is a combination of 10-Acetyl-3,7-dihydroxyphenoxazine (ADHP) (wavelength exc/em = 530-540nm / 590-600nm, a highly sensitive and stable substrate for HRP) and ADHP Dilution Buffer (a stabilized H2O2 solution). Learn more about the fluorogenic substrate, ADHP.
- Tested applicationsSandwich ELISA more details
- Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 96-well PhosphoTracer assay plate (stripwell) 1 unit Adherent plate seal 2 units ADHP (100X) 1 x 120µl ADHP Dilution Buffer 1 x 15ml Assay Control Lysate (lyophilized) 1 x 0.25ml Enhancer Solution 1 x 1ml Lysis Buffer (5X) 1 x 15ml Mouse monoclonal STAT5 (96 assay points) 1 x 3ml Rabbit polyclonal STAT5 (HRP) (96 assay points) 1 x 3ml Stop Solution 1 x 2ml Wash Buffer (10X) 1 x 15ml
- FunctionCarries out a dual function: signal transduction and activation of transcription. Binds to the GAS element and activates PRL-induced transcription.
- Sequence similaritiesBelongs to the transcription factor STAT family.
Contains 1 SH2 domain.
modificationsTyrosine phosphorylated in response to IL-2, IL-3, IL-7, IL-15, GM-CSF, growth hormone, prolactin, erythropoietin and thrombopoietin. Tyrosine phosphorylation is required for DNA-binding activity and dimerization. Serine phosphorylation is also required for maximal transcriptional activity.
- Cellular localizationCytoplasm. Nucleus. Translocated into the nucleus in response to phosphorylation.
- Mammary gland factor
- Signal transducer and activator of transcription 5
- Signal transducer and activator of transcription 5A
- Signal transducer and activator of transcription 5B
- Transcription factor STAT5B
- 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: Q62771 Rat
- SwissProt: P52632 Rat
- Unigene: 437058 Human
Our Abpromise guarantee covers the use of ab119653 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Sandwich ELISA||sELISA. Tested in A431, THP1, NIH3T3|
STAT5 Total PhosphoTracer ELISA Kit images
Using Western blot, a significant stimulation of STAT5 phosphorylation at Tyr694/699 is detected in A431 cells treated with EGF for 10 minutes (+) compared with cells treated with AG1478 (-).
Using the phospho-STAT5 assay kit, a significant stimulation of STAT5 phosphorylation at Tyr694/699 is detected in A431 cells treated with EGF for 10 minutes (+) compared with cells treated with AG1478 (-).
Day 1: A431 cells were seeded at 25K/well in a 96 well tissue culture plate in medium containing 10% FBS. Day 2: The cells were stimulated with various concentrations of EGF for 15 minutes. The media was removed, and the cells were lysed with 125 µL/well freshly prepared Lysis Mix, with shaking for 10 min. Lysates (50 µL) were transferred to a PhosphoTracer assay plate and analyzed for either phospho STAT5 using the standard PhosphoTracer protocol. Briefly, Antibody Mix specific for phospho-STAT5 (50 µL/well), was added to the lysate, and the plate was incubated for 1 hr at room temp, with shaking. The plate was washed and Substrate Mix was added to the wells. The plate was covered in foil and incubated for 10 min with shaking. Signal in the wells was determined using a plate reader.
References for STAT5 Total PhosphoTracer ELISA Kit (ab119653)
ab119653 has not yet been referenced specifically in any publications.