Immunofluorescence analysis of STAT1 [pY701] was done on 70% confluent log phase HeLa cells; untreated and treated (serum starved for 16 hours followed by treatment with 20 ng/ml IFN alpha for 1 hour). The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton™ X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with ABfinity™ STAT1 [pY701] Recombinant Rabbit Oligoclonal Antibody (710078) at 2 µg-4 µg in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 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 with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 Phalloidin (A12381). Panel d is a merged image showing translocated STAT1 [pY701] in nucleus. Panel e shows cytoplasmic localization of STAT1 [pY701] in untreated HeLa cells. Panel f shows competition with STAT1 [pY701] peptide. The images were captured at 20X magnification.
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Recombinant protein corresponding to amino acids 134–218 of human STAT1|
|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 *|
|ChIP assay (ChIP)||3µg|
|Western Blot (WB)||0.5-1µg/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.
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™ oligoclonal antibodies comprise a selection of multiple different recombinant monoclonal antibodies, providing the best of both worlds—the sensitivity of a polyclonal antibody with the specificity of a monoclonal, all delivered with the consistency only found in a recombinant antibody. While functionally the same as a polyclonal antibody—recognizing multiple epitope sites on the target and producing higher detection sensitivity for low abundance targets when compared with monoclonal antibodies—an oligoclonal antibody has a known mixture of light and heavy chains. This exact population can be produced in every lot, circumventing the biological variability typically associated with polyclonal antibody production.
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.
STAT1 (also known as signal transducers and activators of transcription) is a member of the STAT family of transcription factors. STAT proteins undergo phosphorylation in response to cytokines and growth factors and are translocated to the nucleus. STAT1 can be activated by interferon-alpha, interferon-gamma, EGF, PDGF and IL6. The protein is known to regulate several genes which are involved in cell growth, apoptosis, immune responses, and lipid metabolism. The STAT1 gene is located on chromosome 2.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.