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, Mouse, Not Applicable|
|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|
|Immunohistochemistry (Paraffin) (IHC (P))||1:20|
|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.
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.
|Not Applicable||Not Cited||
Consequence of the tumor-associated conversion to cyclin D1b.
PA5-16607 was used in western blot to determine the tumor-associated conversion to cyclin D1b consequences
|Augello MA,Berman-Booty LD,Carr R,Yoshida A,Dean JL,Schiewer MJ,Feng FY,Tomlins SA,Gao E,Koch WJ,Benovic JL,Diehl JA,Knudsen KE||EMBO molecular medicine (7:628)||2015|
Estrogen receptors ß1 and ß2 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|
|Not Applicable||Not Cited||
The p53 inhibitor MDM2 facilitates Sonic Hedgehog-mediated tumorigenesis and influences cerebellar foliation.
PA5-16607 was used in immunohistochemistry to investigate the effect of MDM2 on cerebellar foliation and tumor formation
|Malek R,Matta J,Taylor N,Perry ME,Mendrysa SM||PloS one (6:null)||2011|
Janus kinase 2 is required for the initiation but not maintenance of prolactin-induced mammary cancer.
PA5-16607 was used in immunocytochemistry to explore the contribution of Jak2/Stat5 signaling during mammary cancer initiation and progression
|Sakamoto K,Triplett AA,Schuler LA,Wagner KU||Oncogene (29:5359)||2010|