Immunofluorescence analysis of STAT4 was done on 70% confluent log phase HeLa cells treated with IFN-gamma (50 ng/mL for 20 min). The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton™ X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with STAT4 Rabbit monoclonal Antibody (700185) at 2µg/mL in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor 594 Phalloidin (A12381). Panel d is a merged image showing nuclear localization upon treatment. Panel e shows untreated HeLa cells with cytoplasmic localization. Panel f shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A peptide corresponding to amino acids 730-748 of Q14765.|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||1-5 ug/ml|
|Flow Cytometry (Flow)||1:100-1:500|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:200|
|Western Blot (WB)||3-5 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody is predicted to react with chicken, equine and porcine based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
STAT4 was originally identified using degenerate primers complementary to sequences encoding conserved regions of other STAT proteins. The STAT4 protein is most similar to STAT1 (52%) and STAT3 (47%). Functionally, STAT4 is similar to other STAT family members in that it can be tyrosine phosphorylated by Jak1 or Jak2. STAT4 forms homodimers and heterodimers with related STAT family members. Tyrosine phosphorylated STAT4 can bind the IFN-gamma activated site (GAS). Serine phosphorylation of STAT is also required for maximal transcriptional activity. STAT4 expression is restricted to the thymus, spleen and testis. Until recently the cytokine(s) responsible for activation of STAT4 had not been identified. STAT4 is now known to be activated by the cytokine interleukin 12 (IL-12). IL-12 is required for the T-cell independent induction of IFN-gamma which is a key step in the initial suppression of bacterial and parasitic infections. In addition, IL-12 is required for the development of a Th1 response which is necessary for effective host defense against intracellular pathogens. STAT4-deficient mice display impaired IL-12 development of Th1 cells and enhanced development of Th2 cells. A recent study in mouse has shown that in response to viral infection IFN-a/b activation of STAT4 is required for IFN-g production.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.