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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| Inv, Vrt | ELISA, FC, IP, WB, IH(P) | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | CBL407 |
| Brand Family | Chemicon® |
| Trade Name |
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| Description | Anti-PCNA Antibody, clone PC10 |
| Alternate Names |
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| Background Information | Proliferating cell nuclear antigen (UniProt: P04961; also known as PCNA) is encoded by the Pcna gene (Gene ID: 25737) in rat. PCNA is a homotrimeric nuclear protein that serves as an auxiliary protein of DNA polymerase delta and is involved in the control of eukaryotic DNA replication by increasing the polymerases processability during elongation of the leading strand. It Induces a robust stimulatory effect on the 3-5 exonuclease and 3-phosphodiesterase, but not apurinic-apyrimidinic (AP) endonuclease, APEX2 activities. It is reported to colocalize with CREBBP, EP300 and POLD1 to sites of DNA damage. It acts as a loading platform to recruit DDR proteins that allow completion of DNA replication after DNA damage and promote post-replication repair. PCNA can undergo phosphorylation on tyrosine 211 by EGFR that stabilizes chromatin-associated PCNA. Following DNA damage, it can undergo monoubiquitination to stimulate direct bypass of DNA lesions by specialized DNA polymerases or polyubiquitination to promote recombination-dependent DNA synthesis across DNA lesions by template switching mechanisms. Mutations in PCN gene are known to cause Ataxia-telangiectasia-like disorder 2 (ATLD2) that is characterized by developmental delay, ataxia, hearing loss, and short stature. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
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| Presentation | The monoclonal is presented at a concentration of 100μg/1mL in phosphate buffered saline containing 10mM sodium azide and 1mg/ml bovine serum albumin. We recommend that each laboratory determine an optimum working titre for use in its particular application. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-PCNA Antibody, clone PC10 is a mouse monoclonal antibody for detection of PCNA also known as Proliferating Cell Nuclear Antigen, DNA Polymerase delta Processivity Factor & has been validated in ELISA, FC, IP, WB,IHC(P). |
| Key Applications |
|
| Application Notes | 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 Bouins 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. |
| Biological Information | |
|---|---|
| Immunogen | Rat PCNA induced in the protein A expression vector pR1T2T. |
| Clone | PC10 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| 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. |
| Isotype | IgG2a |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | The protein encoded by this gene is found in the nucleus and is a cofactor of DNA polymerase delta. The encoded protein acts as a homotrimer and helps increase the processivity of leading strand synthesis during DNA replication. In response to DNA damage, this protein is ubiquitinated and is involved in the RAD6-dependent DNA repair pathway. Two transcript variants encoding the same protein have been found for this gene. Pseudogenes of this gene have been described on chromosome 4 and on the X chromosome. |
| Gene Symbol |
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| UniProt Number |
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| UniProt Summary | FUNCTION: SwissProt: P12004 # This protein is an auxiliary protein of DNA polymerase delta and is involved in the control of eukaryotic DNA replication by increasing the polymerases processibility during elongation of the leading strand. SIZE: 261 amino acids; 28769 Da SUBUNIT: Homotrimer. Interacts with KCTD10 (By similarity). Forms a complex with activator 1 heteropentamer in the presence of ATP. Interacts with POLH, POLK, DNMT1, ERCC5/XPG, FEN1, CDC6, APEX2 and POLDIP2. Interacts with EXO1 and SHPRH. SUBCELLULAR LOCATION: Nucleus. PTM: Upon methyl methanesulfonate-induced DNA damage, mono- ubiquitinated by the UBE2B-RAD18 complex on Lys-164. This induces non-canonical poly-ubiquitination on Lys-164 through Lys-63 linkage of ubiquitin moieties by the E2 complex UBE2N-UBE2V2 and the E3 ligase SHPRH, whih is required for DNA repair. DISEASE:SwissProt: P12004 # Antibodies are present in sera from patients with systemic lupus erythematosus. SIMILARITY: SwissProt: P12004 ## Belongs to the PCNA family. |
| Product Usage Statements | |
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| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | For use within 1 month of purchase store at +4°C, for long term storage aliquot antibody into small volumes and store at -20°C. Avoid repeated freeze/ thawing. |
| Packaging Information | |
|---|---|
| Material Size | 100 µg |