Human PCNA ELISA Kit (ab196270)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 0.103 ng/ml
- Range: 0.78 ng/ml - 50 ng/ml
- Sample type: Cell culture extracts, Tissue Extracts
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human PCNA ELISA Kit -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% HeLa extract 8 6.53% Inter-assay Sample n Mean SD CV% HeLa extract 3 10.9% -
Sample type
Cell culture extracts, Tissue Extracts -
Assay type
Sandwich (quantitative) -
Sensitivity
0.103 ng/ml -
Range
0.78 ng/ml - 50 ng/ml -
Recovery
Sample specific recovery Sample type Average % Range Serum 99 89% - 116% Cell culture media 106 103% - 110% Fetal Bovine Serum 111 108% - 115% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
Human PCNA ELISA Kit (ab196270) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of PCNA protein in cell culture extracts and tissue extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Human PCNA with 0.103 ng/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells stripsA 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
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Notes
Proliferating cell nuclear antigen (PCNA) is a 28.7 kDa protein which is an essential component of DNA replication. PCNA protein forms a homotrimeric ring structure which assembles around DNA forming a processivity sliding clamp. Levels of PCNA protein are low in quiescent cells and increase upon mitogen stimulation with peak protein levels localized to the nucleus during S-phase. PCNA interacts with a large number of proteins involved in DNA replication, cell cycle control, and cell cycle check point control.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses. -
Platform
Microplate
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human PCNA Capture Antibody 1 x 600µl 10X Human PCNA Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 5BI 1 x 6ml Human PCNA Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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Function
This protein is an auxiliary protein of DNA polymerase delta and 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. -
Sequence similarities
Belongs to the PCNA family. -
Post-translational
modificationsUpon methyl methanesulfonate-induced DNA damage, mono-ubiquitinated by the UBE2B-RAD18 complex on Lys-164. This induces 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, which is required for DNA repair. 'Lys-63' polyubiquitination prevents genomic instability on DNA damage. 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.
Acetylated in response to UV irradiation. Acetylation disrupts interaction with NUDT15 and promotes degradation. -
Cellular localization
Nucleus. Forms nuclear foci representing sites of ongoing DNA replication and vary in morphology and number during S phase. Together with APEX2, is redistributed in discrete nuclear foci in presence of oxidative DNA damaging agents. - Information by UniProt
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Alternative names
- ATLD2
- cb16
- Cyclin
see all -
Database links
- Entrez Gene: 5111 Human
- Omim: 176740 Human
- SwissProt: P12004 Human
- Unigene: 147433 Human
- Unigene: 728886 Human
Images
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Other - Human PCNA ELISA Kit (ab196270)
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example of PCNA standard curve.Background-subtracted data values (mean +/- SD) are graphed.
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Bar graph displays interpolated values of PCNA of Cell extracts diluted to within the working range of the assay.Background-subtracted data values (mean +/- SD, n = 2) are graphed.
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Titration of HeLa extracts treated with cell cycle arresting compounds and titrated within the working range of the assay.HeLa cells were untreated asynchronous culture and 10 µM Etoposide, 24 hr (S/G2 arrest). PCNA levels were expected to peak during DNA replication in S-phase and return back to normal shortly after exiting S-phase. Background-subtracted data values (mean +/- SD, n = 2) are graphed.
