Overview

  • 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

    PCNA in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit (ab196270) is designed for the quantitative measurement of PCNA protein in Human cell and tissue extracts.


    The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.

  • 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.

  • Tested applications
    Suitable for: Sandwich ELISAmore details
  • Platform
    Microplate

Properties

  • 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
    ab221825 - Antibody Diluent 5BI 1 x 6ml
    Human PCNA Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 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
  • 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
    modifications
    Upon 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
  • Alternative names
    • ATLD2
    • cb16
    • Cyclin
    • DNA polymerase delta auxiliary protein
    • etID36690.10
    • fa28e03
    • fb36g03
    • HGCN8729
    • MGC8367
    • Mutagen-sensitive 209 protein
    • OTTHUMP00000030189
    • OTTHUMP00000030190
    • PCNA
    • Pcna/cyclin
    • PCNA_HUMAN
    • PCNAR
    • Polymerase delta accessory protein
    • Proliferating cell nuclear antigen
    • wu:fa28e03
    • wu:fb36g03
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab196270 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

  • Background-subtracted data values (mean +/- SD) are graphed.

  • Background-subtracted data values (mean +/- SD, n = 2) are graphed.

  • 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.

Protocols

References

This product has been referenced in:
  • Song E  et al. Targeted proteomic assays for quantitation of proteins identified by proteogenomic analysis of ovarian cancer. Sci Data 4:170091 (2017). Read more (PubMed: 28722704) »

See 1 Publication for this product

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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