Recombinant
RabMAb

Recombinant Anti-PCNA antibody [EPR3821] (HRP) (ab193965)

Overview

  • Product name

    Anti-PCNA antibody [EPR3821] (HRP)
    See all PCNA primary antibodies
  • Description

    Rabbit monoclonal [EPR3821] to PCNA (HRP)
  • Host species

    Rabbit
  • Conjugation

    HRP
  • Tested applications

    Suitable for: IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide within Human PCNA aa 100-200. The exact sequence is proprietary.
    Database link: P12004

  • Positive control

    • WB: HeLa, HepG2, HEK293 and A431 whole cell lysates. IHC-P: Normal human colon tissue.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab193965 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
IHC-P 1/10000. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

The use of an HRP/AP polymerized antibody is recommended for a secondary antibody.

WB 1/5000. Detects a band of approximately 29 kDa (predicted molecular weight: 29 kDa).

Target

  • 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
  • Database links

  • Alternative names

    • ATLD2 antibody
    • cb16 antibody
    • Cyclin antibody
    • DNA polymerase delta auxiliary protein antibody
    • etID36690.10 antibody
    • fa28e03 antibody
    • fb36g03 antibody
    • HGCN8729 antibody
    • MGC8367 antibody
    • Mutagen-sensitive 209 protein antibody
    • OTTHUMP00000030189 antibody
    • OTTHUMP00000030190 antibody
    • PCNA antibody
    • Pcna/cyclin antibody
    • PCNA_HUMAN antibody
    • PCNAR antibody
    • Polymerase delta accessory protein antibody
    • Proliferating cell nuclear antigen antibody
    • wu:fa28e03 antibody
    • wu:fb36g03 antibody
    see all

Images

  • IHC image of PCNA staining in a section of formalin-fixed paraffin-embedded normal human colon tissue*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab193965 at 1/100 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • All lanes : Anti-PCNA antibody [EPR3821] (HRP) (ab193965) at 1/5000 dilution

    Lane 1 : HeLa whole cell lysate (ab150035)
    Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 4 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 29 kDa
    Observed band size: 29 kDa


    Exposure time: 30 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab193965 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

References

ab193965 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (Lung and Breast)
Gel Running Conditions
Reduced Denaturing
Loading amount
30 µg
Specification
Lung and Breast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Dec 19 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Skin and tongue basal epithelial cells, Molar root)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA pH=8
Permeabilization
Yes - 0.05% tween20
Specification
Skin and tongue basal epithelial cells, Molar root
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde

Dr. Juan Fons Romero

Verified customer

Submitted Jun 28 2018

Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Skin, Palate and Tongue epithelia, Brain (Neural S)
Antigen retrieval step
None
Permeabilization
Yes - 0.05% tween20
Specification
Skin, Palate and Tongue epithelia, Brain (Neural S
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde

Dr. Juan Fons Romero

Verified customer

Submitted Jun 28 2018

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