Phospho-STAT5a (Y694) ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-STAT5a (Y694) in Human in Cell Lysate samples.
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Phospho-STAT5a (Y694) ELISA Kit is a Semi-quantitative ELISA kit for the measurement of Phospho-STAT5a (Y694) in Human in Cell Lysate samples.
Phospho-STAT5a (Y694) ELISA Kit (ab279943) 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. 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. The wells are washed and rabbit anti-phospho-STAT5a (Y694) antibody is used to detect phosphorylated STAT5a. After washing away unbound antibody, HRP conjugated anti-rabbit IgG 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) bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
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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).
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.
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.
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.
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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.
A431 cells treated with/without EGF.
A431 cells were treated or untreated with EGF.
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