PA5-16227

antibody from Invitrogen Antibodies

Targeting: CCNC CycC

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Data presented on provider website)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Data presented on provider website)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-16227

Provider

Invitrogen Antibodies

Product name

Anti-Cyclin C Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

PA5-16227 targets Cyclin C in immunofluorescence, immunoprecipitation, and Western blot applications and shows reactivity with Canine, Human, mouse, and Rat samples. The PA5-16227 immunogen is a synthetic peptide corresponding to aa 290-303 (Cys-QGPNGSQNSSYSQS) from the C-terminal of human cyclin C.

Reactivity

Human, Mouse, Rat, Canine

Host

Rabbit

Isotype

IgG

Vial size

500 µL

Concentration

1 mg/mL

Storage

4° C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on nuclear enriched extracts of HeLa (Lane 1), Hep G2 (Lane 2) and NIH/3T3 (Lane 3). The blots were probed with Anti- Cyclin C Rabbit Polyclonal Antibody (Product # PA5-16227, 2 µg/mL) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution. A ~ 33 kDa band corresponding to Cyclin C was observed across all cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow Cytometry analysis of Cyclin C was done on HeLa cells (untreated, red histogram) and HeLa cells serum starved and serum released for 6 hours (blue histogram). Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Cyclin C Rabbit Polyclonal Antibody (PA5-16227) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10, 000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells.