| Applications: |
CLIA |
| Reactivity: |
Human |
| Note: |
FOR SCIENTIFIC EDUCATIONAL RESEARCH USE ONLY (RUO). MUST NOT BE USED IN DIAGNOSTIC OR OTHER MEDICAL APPLICATIONS. |
| Sensitivity: |
37.5pg/mL |
| Detection Limit: |
62.5~4000pg/mL |
| Short Description: |
This human PCNA kit is a highly sensitive in-vitro chemiluminescent immunoassay for the measurement of trace amounts of analytes. |
| Storage Instruction: |
If unopened the kit may be stored at 2-8°C for up to 1 month. If the kit will not be used within 1 month, store the components separately, according to the component table in the manual. |
| Assay Time: |
3.5h |
| Detection: |
Chemiluminescence |
| Gene Symbol: |
PCNA |
| Gene ID: |
5111 |
| Uniprot ID: |
PCNA_HUMAN |
| Specificity: |
This kit recognizes Human PCNA in samples. No significant cross-reactivity or interference between Human PCNA and analogues was observed. |
| Sample Type: |
Serum, plasma and other biological fluids |
| Post Translational Modifications | Phosphorylated. Phosphorylation at Tyr-211 by EGFR stabilizes chromatin-associated PCNA. Acetylated by CREBBP and p300/EP300.preferentially acetylated by CREBBP on Lys-80, Lys-13 and Lys-14 and on Lys-77 by p300/EP300 upon loading on chromatin in response to UV irradiation. Lysine acetylation disrupts association with chromatin, hence promoting PCNA ubiquitination and proteasomal degradation in response to UV damage in a CREBBP- and EP300-dependent manner. Acetylation disrupts interaction with NUDT15 and promotes degradation. Ubiquitinated. Following DNA damage, can be either monoubiquitinated to stimulate direct bypass of DNA lesions by specialized DNA polymerases or polyubiquitinated to promote recombination-dependent DNA synthesis across DNA lesions by template switching mechanisms. Following induction of replication stress, monoubiquitinated by the UBE2B-RAD18 complex on Lys-164, leading to recruit translesion (TLS) polymerases, which are able to synthesize across DNA lesions in a potentially error-prone manner. An error-free pathway also exists and requires non-canonical polyubiquitination on Lys-164 through 'Lys-63' linkage of ubiquitin moieties by the E2 complex UBE2N-UBE2V2 and the E3 ligases, HLTF, RNF8 and SHPRH. This error-free pathway, also known as template switching, employs recombination mechanisms to synthesize across the lesion, using as a template the undamaged, newly synthesized strand of the sister chromatid. Monoubiquitination at Lys-164 also takes place in undamaged proliferating cells, and is mediated by the DCX(DTL) complex, leading to enhance PCNA-dependent translesion DNA synthesis. Sumoylated during S phase. Methylated on glutamate residues by ARMT1/C6orf211. |
| Function | Auxiliary protein of DNA polymerase delta and epsilon, is involved in the control of eukaryotic DNA replication by increasing the polymerase's processibility during elongation of the leading strand. Induces a robust stimulatory effect on the 3'-5' exonuclease and 3'-phosphodiesterase, but not apurinic-apyrimidinic (AP) endonuclease, APEX2 activities. Has to be loaded onto DNA in order to be able to stimulate APEX2. Plays a key role in DNA damage response (DDR) by being conveniently positioned at the replication fork to coordinate DNA replication with DNA repair and DNA damage tolerance pathways. Acts as a loading platform to recruit DDR proteins that allow completion of DNA replication after DNA damage and promote postreplication repair: Monoubiquitinated PCNA leads to recruitment of translesion (TLS) polymerases, while 'Lys-63'-linked polyubiquitination of PCNA is involved in error-free pathway and employs recombination mechanisms to synthesize across the lesion. |
| Protein Name | Proliferating Cell Nuclear AntigenPcnaCyclin |
| Database Links | Reactome: R-HSA-110312Reactome: R-HSA-110314Reactome: R-HSA-110320Reactome: R-HSA-1362277Reactome: R-HSA-174411Reactome: R-HSA-174414Reactome: R-HSA-174417Reactome: R-HSA-174437Reactome: R-HSA-4615885Reactome: R-HSA-5358565Reactome: R-HSA-5358606Reactome: R-HSA-5651801Reactome: R-HSA-5655862Reactome: R-HSA-5656121Reactome: R-HSA-5656169Reactome: R-HSA-5685942Reactome: R-HSA-5696397Reactome: R-HSA-5696400Reactome: R-HSA-6782135Reactome: R-HSA-6782210Reactome: R-HSA-6804114Reactome: R-HSA-69091Reactome: R-HSA-69166Reactome: R-HSA-69183Reactome: R-HSA-69205Reactome: R-HSA-8866654 |
| Cellular Localisation | NucleusColocalizes With CrebbpEp300 And Pold1 To Sites Of Dna DamageForms Nuclear Foci Representing Sites Of Ongoing Dna Replication And Vary In Morphology And Number During S PhaseCo-Localizes With Smarca5/Snf2h And Baz1b/Wstf At Replication Foci During S PhaseTogether With Apex2Is Redistributed In Discrete Nuclear Foci In Presence Of Oxidative Dna Damaging Agents |
| Alternative CLIA Names | Proliferating Cell Nuclear Antigen CLIA kitPcna CLIA kitCyclin CLIA kitPCNA CLIA kit |
| Specificity | This kit recognizes Human PCNA in samples. No significant cross-reactivity or interference between Human PCNA and analogues was observed. |
| Reproducibility | Both intra-CV and inter-CV are |
Information sourced from Uniprot.org
| Item | Specifications | Storage |
| Micro CLIA Plate (Dismountable) | 96T: 8 wells ×12 strips strips | -20℃, 6 months |
| Reference Standard | 96T: 2 vials 48T: 1 vial | -20℃, 6 months |
| Concentrated Biotinylated Detection Ab (100×) | 96T: 1 vial, 120 μL 60 μL | -20℃, 6 months |
| Concentrated HRP Conjugate (100×) | 96T: 1 vial, 120 μL 60 μL | -20℃ (Protect from light), 6 months |
| Reference Standard & Sample Diluent | 1 vial, 20 mL | 2-8°C, 6 months |
| Biotinylated Detection Ab Diluent | 1 vial, 14 mL | 2-8°C, 6 months |
| HRP Conjugate Diluent | 1 vial, 14 mL | 2-8°C, 6 months |
| Concentrated Wash Buffer (25×) | 1 vial, 30 mL | 2-8°C, 6 months |
| Substrate Reagent A | 1 vial, 5 mL | 2-8℃ (Protect from light) |
| Substrate Reagent B | 1 vial, 5 mL | 2-8℃ (Protect from light) |
| Plate Sealer | 5 pieces | |
| Manual | 1 copy | |
| Certificate of Analysis | 1 copy | |
| Sample Type | Range (%) | Average Recovery (%) |
| Serum(n=8) | 97-109 | 103 |
| EDTA plasma(n=8) | 90-108 | 98 |
| Cell culture media(n=8) | 84-97 | 91 |
| | Intra-assay Precision | Intra-assay Precision | Intra-assay Precision | Inter-assay Precision | Inter-assay Precision | Inter-assay Precision |
| Sample | 1.00 | 2.00 | 3.00 | 1.00 | 2.00 | 3.00 |
| n | 20.00 | 20.00 | 20.00 | 20.00 | 20.00 | 20.00 |
| Mean (pg/mL) | 216.97 | 430.99 | 1672.32 | 207.91 | 412.45 | 1759.76 |
| Standard deviation | 25.99 | 47.67 | 105.02 | 25.37 | 45.78 | 113.86 |
| CV (%) | 11.98 | 11.06 | 6.28 | 12.20 | 11.10 | 6.47 |
12 months for antibodies. 6 months for ELISA Kits. Please see website T&Cs for further guidance
