Immunofluorescence analysis of STAT5 beta was done on 70% confluent log phase HeLa cells. 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 STAT5 beta Mouse Monoclonal Antibody (135300) at 2 µg/mL and incubated for 3 hours at room temperature and then labeled with Alexa Flour 488 Rabbit Anti-Mouse IgG Secondary Antibody (A11059) 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 cytoplasmic localization. Panel e shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Rat, Human, Mouse|
|Published species reactivity||Virus, Bovine, Human, Mouse, Not Applicable|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||A peptide corresponding to the unique carboxy-terminus of murine STAT5b coupled to a protein carrier.|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|ChIP assay (ChIP)||5 ug/ml|
|ELISA (ELISA)||Assay Dependent|
|Gel Shift (GS)||Assay Dependent|
|Immunocytochemistry (ICC)||1-3 ug/ml|
|Immunofluorescence (IF)||1-3 ug/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20-1:200|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1-3 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.
The protein encoded by this gene 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. This protein mediates the signal transduction triggered by various cell ligands, such as IL2, IL4, CSF1, and different growth hormones. It has been shown to be involved in diverse biological processes, such as TCR signaling, apoptosis, adult mammary gland development, and sexual dimorphism of liver gene expression. This gene was found to fuse to retinoic acid receptor-alpha (RARA) gene in a small subset of acute promyelocytic leukemias (APLL). The dysregulation of the signaling pathways mediated by this protein may be the cause of the APLL.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
DNA methylation and transcription in a distal region upstream from the bovine AlphaS1 casein gene after once or twice daily milking.
13-5300 was used in EMSA to investigate the role of CpG methylation in milk production
|Nguyen M,Boutinaud M,Pétridou B,Gabory A,Pannetier M,Chat S,Bouet S,Jouneau L,Jaffrezic F,Laloë D,Klopp C,Brun N,Kress C,Jammes H,Charlier M,Devinoy E||PloS one (9:null)||2014|
Identification of STAT5A and STAT5B target genes in human T cells.
13-5300 was used in ChIP-seq, immunocytochemistry, and western blot to investigate genes regulated by STAT5A and/or STAT5B in human CD4(+) T cells.
|Kanai T,Seki S,Jenks JA,Kohli A,Kawli T,Martin DP,Snyder M,Bacchetta R,Nadeau KC||PloS one (9:null)||2014|
Prolactin signalling in the mouse hypothalamus is primarily mediated by signal transducer and activator of transcription factor 5b but not 5a.
13-5300 was used in immunohistochemistry - free floating to report that STAT5b and not STAT5a is the primary mediator prolactin signaling in the hypothalamus.
|Yip SH,Eguchi R,Grattan DR,Bunn SJ||Journal of neuroendocrinology (24:1484)||2012|
|Not Applicable||Not Cited||
Identification of human STAT5-dependent gene regulatory elements based on interspecies homology.
13-5300 was used in immunoprecipitation and western blot to identify NCAM1 as a novel STAT5 target
|Nelson EA,Walker SR,Li W,Liu XS,Frank DA||The Journal of biological chemistry (281:26216)||2006|
|Not Applicable||Not Cited||
Isolation of unique STAT5 targets by chromatin immunoprecipitation-based gene identification.
13-5300 was used in western blot to investigate STAT5-regulated genes
|Nelson EA,Walker SR,Alvarez JV,Frank DA||The Journal of biological chemistry (279:54724)||2004|
|Activated signal transducer and activator of transcription (STAT) 3: localization in focal adhesions and function in ovarian cancer cell motility.||Silver DL,Naora H,Liu J,Cheng W,Montell DJ||Cancer research (64:3550)||2004|
|Mouse||Not Cited||Suppressor of cytokine signaling-1 is a critical regulator of interleukin-7-dependent CD8+ T cell differentiation.||Chong MM,Cornish AL,Darwiche R,Stanley EG,Purton JF,Godfrey DI,Hilton DJ,Starr R,Alexander WS,Kay TW||Immunity (18:475)||2003|
Stat5a is tyrosine phosphorylated and nuclear localized in a high proportion of human breast cancers.
13-5300 was used in immunoprecipitation and western blot to examine the expression, tyrosine phosphorylation status and nuclear localization of Stat5a in human breast cancers.
|Cotarla I,Ren S,Zhang Y,Gehan E,Singh B,Furth PA||International journal of cancer (108:665)||2004|