Technical guide & protocol Tech Support

Product Specifications

Analytical sensitivity

<0.9 U/mL

Assay range

1.57-100 U/mL

Sample type/volume

Cell Lysate
10 µL

Hands-on time

1 hr 20 min

Time-to-result

4 hr

Homogenous (no wash)

No

Interassay CV

4.5%

Intraassay CV

3.9%

Instrument

Colorimetric Microplate Reader

Product size

96 Tests

Contents

Pre-coated 96 well plate
Standard
Standard Dilution Buffer
Detection Antibody
anti-rabbit-HRP
HRP Diluent
Wash Buffer
Chromogen
Stop Solution
Adhesive Plate Covers

Shipping conditions

Wet ice

Storage

2-8°C

Protein name

STAT3

Target site-specificity

Phosphorylated Tyr705

Protein family

Transcription Factors

Species (tested)

Human, Mouse, Rat

Assay kit format

Sandwich ELISA Kit

Detector antibody conjugate

HRP

Label or dye

HRP

About This Kit

The Human STAT3 (Phospho) [pY705] ELISA Kit is a solid-phase sandwich Enzyme-Linked Immunosorbent Assay (ELISA) designed to detect and quantify the level of STAT3 (Phospho) [pY705] in fresh or frozen human cell lysates. Cross-reactivity has been observed in mouse and rat cells. The assay recognizes both natural and recombinant human STAT3 (Phospho) [pY705].

Principle of the method

A monoclonal capture antibody specific for STAT3 (Phospho) [pY705]) has been coated onto the wells of the 96-well plate. During the first incubation, standards of known content and unknown samples are pipetted into the wells and the antigen binds to the immobilized (capture) antibody. After washing, a rabbit antibody specific for the target protein is added to the wells and serves as a detection antibody by binding to the immobilized protein captured during the first incubation. After washing, a horseradish peroxidase labeled anti-rabbit IgG is added. This binds to the detection antibody to complete the four member sandwich. After a third incubation and washing to remove all the unbound enzyme, a substrate solution (TMB) is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of target protein present in the original specimen and the optical density can be read on a standard microplate reader.

Rigorous validation

Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

Target information

STAT3 belongs to the family of STAT (signal transducers and activators of transcription) proteins which are phosphorylated by receptor associated kinases, translocate to the nucleus, and act as transcription factors in response to cytokines and growth factors. Coactivators such as CREB-binding protein are required for the transcriptional activation by STAT3. STAT3 can also be activated by Interferon-alpha, Interferon-gamma, EGF, PDGF and IL6. Phosphorylation on tyrosine 705 by JAK1 and JAK2 is essential for STAT3 dimer formation, nuclear translocation, and DNA binding activity. In humans, the STAT3 gene is located on the q arm of chromosome 17. STAT3 has been shown to be activated by IFN-alpha but not IFN-beta. The transcription factors associated with STAT3 are c-Jun and cyclic AMP-responsive enhancer binding protein (CREB). STAT3 mediates the expression of a variety of genes in response to cell stimuli, and thus plays a key role in many cellular processes such as cell growth and apoptosis. The small GTPase Rac1 has been shown to bind and regulate the activity of STAT3 while the PIAS3 protein is a specific inhibitor of STAT3. Three alternatively spliced transcript variants encoding distinct isoforms of STAT3 have been described. Deletion of the STAT3 gene in knock-out mice was lethal at the early embryonic stage.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Gene aliases : 1110034C02Rik, ADMIO, ADMIO1, APRF, AW109958, HIES, STAT3

Gene ID : (Human) 6774, (Rat) 25125, (Mouse) 20848

Gene symbol : STAT3, Stat3, Stat3

Protein Aliases : acute phase response factor, Acute-phase response factor, DNA-binding protein APRF, Signal transducer and activator of transcription 3, signal transducer and activator of transcription 3 (acute-phase response factor)

UniProt ID (Human) P40763, (Rat) P52631, (Mouse) P42227

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