Immunofluorescence analysis of Phospho-Retinoblastoma pSer608 was done on 70% confluent log phase Colo cells treated with serum starvation for 16 hours. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Phospho-Retinoblastoma pSer608 Antibody (51B7) Mouse Monoclonal Antibody (MA512584) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) at a dilution of 1:2000 for 45 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® 555 Rhodamine Phalloidin (Product # R415, 1:300).Panel d is a merged image showing punctate nuclear localization. Panel e is untreated cell with no signal. Panel f is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Rat, Mouse, Human, Not Applicable|
|Host / Isotype||Mouse / IgG1|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Western Blot (WB)||1-2 µg/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 was orginally validated as part of a Thermo Scientific Cellomics High Content Screening Kit. The antibody sold separately may have slightly different performance and may need to be further optimized for the best results.
Rb is a tumor suppressor nuclear phosphoprotein capable of binding to DNA. It is phosphorylated on serine and threonine, but not on tyrosine residues. It forms a complex with SV40 large T antigen, adenovirus E1A, and human papilloma virus-16 E. Rb protein may act by regulating transcription and loss of its function leads to uncontrolled cell growth. Aberrations in the RB gene have been implicated in cancers of breast, colon, prostate, kidney, nasopharynx, and leukemia. Phosphorylation of Rb at S608 depends on CDK4.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
MuRF1 mono-ubiquitinates TR¿ to inhibit T3-induced cardiac hypertrophy in vivo.
MA5-12584 was used in western blot to assess inhibition of T3-induced cardiac hypertrophy in vivo by MuRF1 mono-ubiquitinating TR-alpha
|Wadosky KM,Berthiaume JM,Tang W,Zungu M,Portman MA,Gerdes AM,Willis MS||Journal of molecular endocrinology (56:273)||2016|
Selective inhibition of rRNA transcription downregulates E2F-1: a new p53-independent mechanism linking cell growth to cell proliferation.
MA5-12584 was used in western blot to investigate the influence of selective inhibition of rRNA transcription on the cell cycle of human cancer cells
|Donati G,Brighenti E,Vici M,Mazzini G,Treré D,Montanaro L,Derenzini M||Journal of cell science (124:3017)||2011|
Bexarotene induces cellular senescence in MMTV-Neu mouse model of mammary carcinogenesis.
MA5-12584 was used in immunohistochemistry to study the role of cellular senescence in the beneficial effects of bexarotene in a murine mammary carcinogenesis model
|Shilkaitis A,Bratescu L,Green A,Yamada T,Christov K||Cancer prevention research (Philadelphia, Pa.) (6:299)||2013|
UM-SCC-104: a new human papillomavirus-16-positive cancer stem cell-containing head and neck squamous cell carcinoma cell line.
MA5-12584 was used in immunohistochemistry to develop a novel HPV-16-positive head and neck carcinima cell line containing a significant proportion of cancer stem cells
|Tang AL,Hauff SJ,Owen JH,Graham MP,Czerwinski MJ,Park JJ,Walline H,Papagerakis S,Stoerker J,McHugh JB,Chepeha DB,Bradford CR,Carey TE,Prince ME||Head and neck (34:1480)||2012|
Proliferation capacity of the renal proximal tubule involves the bulk of differentiated epithelial cells.
MA5-12584 was used in immunohistochemistry to study the role of undifferentiated epithelial cells in the proliferation capacity of the renal proximal tubule
|Vogetseder A,Picard N,Gaspert A,Walch M,Kaissling B,Le Hir M||American journal of physiology. Cell physiology (294:C22)||2008|