The Human STAT5 alpha (Total) ELISA Kit is a solid-phase sandwich Enzyme-Linked Immunosorbent Assay (ELISA) designed to detect and quantify the level of STAT5 alpha (Total) 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 STAT5 alpha (Total), independent of its phosphorylation state.
Principle of the method
A monoclonal capture antibody specific for human STAT5 alpha (Total) 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.
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.
STAT5 alpha is a member of the STAT family of transcription factors. In response to cytokines and growth factors, STAT family members are phosphorylated by the receptor associated kinases, and then form homo- or heterodimers that translocate to the cell nucleus where they act as transcription activators. STAT5 alpha is activated by, and mediates the responses of many cell ligands, such as IL2, IL3, IL7 GM-CSF, erythropoietin, thrombopoietin, and different growth hormones. Activation of STAT5 alpha in myeloma and lymphoma associated with a TEL/JAK2 gene fusion is independent of cell stimulus and has been shown to be essential for the tumorigenesis. The mouse counterpart of STAT5 alpha is found to induce the expression of BCL2L1/BCL-X(L), which suggests the antiapoptotic function of this protein in cells. STAT5 alpha, along with STAT5 beta, were identified in mouse. The amino acid sequences of STAT5 alpha and STAT5 beta show 96% sequence similarity, and both proteins are co expressed in most tissues in virgin and lactating mice. However, differential accumulation of STAT5 alpha and STAT5 beta mRNA has been reported for both muscle and mammary tissue. STAT5 alpha is critically involved in a variety of physiological functions, including reproduction, lactation, immune function and somatic growth.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.