Analysis Note

ControlRat kidney, Human tonsil, lymph node tissue

Application

Research CategoryEpigenetics & Nuclear Function

Use Anti-PCNA Antibody, clone PC10 (mouse monoclonal antibody) validated in ELISA, FC,IHC(P), IP, WB to detect PCNA also known as Proliferating Cell Nuclear Antigen, DNA Polymerase delta Processivity Factor.

Western blot: 1:250-1:1000. Nuclear extracts are preferred over whole cell protein extracts.

Immunohistochemistry: 1:20-1:200. Recommended for paraffin embedded tissue sections only. May be used on material fixed in a wide range of fixatives including formalin (buffered and unbuffered), methacarn and Bouin′s reagent. The time of fixation can markedly affect the intensity of PCNA immunoreactivity. Staining is reduced (and may be abolished) if sections are baked onto glass slides. Air-drying overnight on poly-L-lysine coated slides is recommended. 60 minute incubation at 25°C with standard ABC technique is recommended.

Immunoprecipitation

Indirect Flow Cytometry

Optimal working dilutions must be determined by the end user.

Research Sub CategoryCell Cycle, DNA Replication & Repair

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

Proliferating cell nuclear antigen (PCNA) is a 36 kDa molecule that is highly conserved between species. PCNA was first identified as the antigen for a subpopulation of autoantibodies in patients with systemic lupus erythematosus (Miyachi, 1978; Takasaki, 1984; Ogata, 1987). It has since been determined that PCNA serves as a co-factor for DNA polymerase delta in S-phase as well as during DNA synthesis associated with mechanisms involved in DNA damage repair (Tan, 1987; Bravo, 1987). The temporal specificity of PCNA expression makes it an ideal marker for cell proliferation. PCNA begins to accumulate during the G1 phase of the cell cycle, is most abundant during the S phase, and declines during the G2/M phase (Kurki, 1988). Since the half-life of PCNA exceeds 20 hours, it may be possible to detect the protein in non-cycling cells.

Immunogen

Rat PCNA made in the protein A vector pR1T2T.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Protein A purified

Format: Purified

Protein A Purified mouse immunoglobulin in PBS with 0.1% sodium azide as a preservative.

Quality

Tested

Specificity

Clone PC10 recognizes PCNA from all vertebrate and insect species tested and has been used to identify transformed cells (Kurki, 1988), proliferating cells in solid tumors (Smetana, 1983), and blast cells in leukemia patients (Takasaki, 1984). Immunohistochemistry with clone PC10 has been used to study the expression of PCNA in paraffin sections of normal tissues and lymphoid neoplasms (Hall, 1990). In proliferating cells, PCNA staining pattern is predominantly nuclear. By western blot, the antibody detects a polypeptide migrating at 36 kDa with an isoelectric point of 4.8.

Storage and Stability

Maintain for 1 year at 2–8°C from date of shipment. Aliquot to avoid repeated freezing and thawing. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Target description

36 kDa