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Immunofluorescence analysis of Cyclin D1 / Bcl-1 was performed using 70% confluent log phase MCF-7 cells. 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 Cyclin D1 / Bcl-1 Rabbit Polyclonal Antibody (Product # PA5-16607) at 2µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) 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 (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Rat|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide derived from the C-terminal of human cyclin D1|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunoprecipitation (IP)||10 µg/ml|
|Western Blot (WB)||4-6 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.
|Western Blot (WB)||See 1 publications below|
PA5-16607 targets Cyclin D1/Bcl-1 in immunofluorescence, immunoprecipitation, and Western blot applications and shows reactivity with mouse, Rat, and Human samples.
The PA5-16607 immunogen is a synthetic peptide derived from the C-terminal of human cyclin D1.
Cyclin D1or PRAD-1 or bcl-1 is one of the key cell cycle regulators, and functions in association with cdk4 and/or cdk6 by phosphorylating the Rb protein. It is a putative proto-oncogene overexpressed in a wide variety of human neoplasms including mantle cell lymphomas (MCL).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Estrogen receptors β1 and β2 have opposing roles in regulating proliferation and bone metastasis genes in the prostate cancer cell line PC3.
PA5-16607 was used in western blot to study the tumor suppressing activity of ER-beta1 and the tumor promoting activity of ER-beta2 in prostate cancer cells
|Dey P,Jonsson P,Hartman J,Williams C,Ström A,Gustafsson JÅ||Molecular endocrinology (Baltimore, Md.) (26:1991)||2012|
B-cell CLL/lymphoma 1; B-cell lymphoma 1 protein; BCL-1 oncogene; G1/S-specific cyclin D1; G1/S-specific cyclin-D1; PRAD1 oncogene
AI327039; bcl-1; BCL1; CCND1; cD1; CycD1; Cyl-1; D11S287E; PRAD1; U21B31