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Phospho-STAT5a (Y694) and Total STAT5a ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-STAT5a (Y694) and Total STAT5a in Human in Cell Lysate samples.

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Key facts

Detection method

Colorimetric

Sample types

Cell Lysate

Assay type

Semi-quantitative

Reactive species

Human

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Reactivity data

Application

ELISA

Reactivity

Reacts

Dilution info

-

Notes

-

What's included?

1 x 96 Tests
Components
100X HRP-Streptavidin Concentrate
1 x 200 µL
20X Wash Buffer
1 x 25 mL
2X Cell lysate buffer
1 x 5 mL
5X Assay Diluent
1 x 15 mL
Biotinylated anti-pan-STAT5a detection antibody
1 x 1 Vial
HRP-conjugated anti-rabbit IgG concentrate (500X)
1 x 25 µL
Pan-STAT5a Coated Microplate
1 x 1 Unit
Positive Control - A431 cell lysate
1 x 1 Vial
Rabbit anti-phospho-STAT5a (Y694)-antibody
1 x 1 Vial
Stop Solution
1 x 8 mL
TMB One-Step Substrate Reagent
1 x 12 mL

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Phospho-STAT5a (Y694) and Total STAT5a ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-STAT5a (Y694) and Total STAT5a in Human in Cell Lysate samples.

Key facts

Detection method

Colorimetric

Sample types

Cell Lysate

Assay type

Semi-quantitative

Reactive species

Human

Assay Platform

Pre-coated microplate (12 x 8 well strips)

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

Storage information

-20°C

Notes

Phospho-STAT5a (Y694) and Total STAT5a ELISA Kit (ab279944) is a very rapid, convenient and sensitive assay kit that can monitor the activation or function of important biological pathways in human cell lysates. By determining phosphorylated STAT5a protein in your experimental model system, you can verify pathway activation in your cell lysates. You can simultaneously measure numerous different cell lysates without spending excess time and effort in performing a Western Blotting analysis.

This Sandwich ELISA kit is an in vitro enzyme-linked immunosorbent assay for the measurement of human phospho-STAT5a and total STAT5a. An anti-pan STAT5a antibody has been coated onto a 96-well plate. Samples are pipetted into the wells and STAT5a present in a sample is bound to the wells by the immobilized antibody and the wells are washed. In select wells, rabbit anti-phospho STAT5a (Y694) antibody is added to detect phosphorylated STAT5a. In the remaining wells, biotinylated anti-pan-STAT5a antibody is used to detect pan STAT5a. After washing away unbound antibody, HRP-conjugated anti-rabbit IgG or HRP-Streptavidin is pipetted into the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of STAT5a (Y694) or pan STAT5a bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.

Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

Supplementary info

Biological summary

Many systems use STAT5a to regulate cell proliferation differentiation and survival. It often forms a complex with other proteins like STAT5b which is a closely related counterpart and both contribute to similar cellular processes. STAT5a can dimerize upon activation allowing it to move to the cell nucleus to influence gene expression directly.

Mechanical summary

STAT5a also known as Signal Transducer and Activator of Transcription 5A acts as a transcription factor in the cell. STAT5a weighs approximately 90 kDa and expresses in various tissues including the liver muscle and hematopoietic cells. It plays a central role in mediating cellular responses to cytokines and growth factors particularly those in the interleukin-2 (IL-2) family and growth hormone (GH).

Pathway

Researchers place STAT5a prominently in the JAK-STAT signaling pathway. This pathway is essential for transmitting information received from extracellular chemical signals to the cell nucleus resulting in the activation of genes controlling cellular functions. STAT5a works together with Janus kinases (JAKs) to mediate various biological responses including hormone signaling and immune function. It also interacts with proteins like IL-2 receptor subunits which are involved in many cytokine-driven processes.

Disease

Disturbances in STAT5a activity link specifically with conditions like leukemia and breast cancer. Overactivation or mutation can lead to uncontrolled cell growth and survival contributing to these diseases. In leukemia STAT5a frequently partners with other proteins like BCR-ABL which is known for its role in chronic myeloid leukemia influencing oncogenic pathways. In breast cancer alterations in STAT5a signaling may interact with estrogen and progesterone receptors affecting tumor progression and response to therapies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

2 product images

  • Sandwich ELISA - Phospho-STAT5a (Y694) and Total STAT5a ELISA Kit (ab279944), expandable thumbnail

    Sandwich ELISA - Phospho-STAT5a (Y694) and Total STAT5a ELISA Kit (ab279944)

    Positive Control.

    A431 cells were treated with recombinant human EGF at 37°C for 20 min.

    Cells were solubilzed at 4 x 107 cells/ml in lysis buffer.

    Serial dilutions of lysates were analyzed in this ELISA.

  • Sandwich ELISA - Phospho-STAT5a (Y694) and Total STAT5a ELISA Kit (ab279944), expandable thumbnail

    Sandwich ELISA - Phospho-STAT5a (Y694) and Total STAT5a ELISA Kit (ab279944)

    A431 cells treated with/without EGF.

    A431 cells were treated with recombinant human EGF at 37°C for 20 min.

    Cells were solubilzed at 4 x 107 cells/ml in lysis buffer.

    Serial dilutions of lysates were analyzed in this ELISA.

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com

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