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Proliferating Cell Nuclear Antigen Peptide 3

Antibody: CPTC-PCNA-3
Background

Catalog Number:

CPTC-PCNA-3

RRID:

AB_2888965

Target Antigen:

Proliferating Cell Nuclear Antigen Peptide 3

Isotype:

IgG

Species:

Rabbit Monoclonal Antibody

Last Updated:

05/17/2024

Antigen Recognition(s):

Peptide, Endogenous

Thematic Panel(s):

DNA Damage Response (DDR) Pathway

External Links
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Characterization Data [Compare Characterization Data]
  Affinity Measurement by SPR
Click to enlarge image Affinity and binding kinetics of CPTC-PCNA-3 and full-length PCNA recombinant protein were measured using surface plasmon resonance. CPTC-PCNA-3 antibody was captured on protein A biosensor chip. PCNA recombinant protein at 1024 nM, 256 nM, 64 nM, 16 nM, and 4 nM was used as analyte. All binding data were double referenced and analyzed globally using a 1:1 fitting model. Kinetic constant kd is outside the limits that can be measured by the instrument. Kinetic constants cannot be uniquely determined. Try to extend dissociation time. Click image to enlarge

CPTC-PCNA-3 Affinity and Kinetics (Surface Plasmon Resonance)

Result: Positive

Affinity and binding kinetics of CPTC-PCNA-3 and full-length PCNA recombinant protein were measured using surface plasmon resonance. CPTC-PCNA-3 antibody was captured on protein A biosensor chip. PCNA recombinant protein at 1024 nM, 256 nM, 64 nM, 16 nM, and 4 nM was used as analyte. All binding data were double referenced and analyzed globally using a 1:1 fitting model. Kinetic constant kd is outside the limits that can be measured by the instrument. Kinetic constants cannot be uniquely determined. Try to extend dissociation time.


Characterization SOP Files

  IHC HPA
Download This PDF contains the evaluation results provided by the Human Protein Atlas (www.proteinatlas.org) (164.1 KB)

CPTC-PCNA-3 Evaluation by the Human Protein Atlas

Result: Positive

This PDF contains the evaluation results provided by the Human Protein Atlas (www.proteinatlas.org)


Characterization SOP Files

  IHC Tissue

CPTC-PCNA-3 IHC Tissue

Result: Negative

This antibody is not suitable for use in an Immunohistochemistry format as described in SOP M-106.


Characterization SOP Files

  IP
Click to enlarge image Immuno-precipitation performed using CPTC-PCNA-3 as capture antibody against the over-expressed lysate of PCNA (Myc-tagged). Eluates were tested in automated western blot using the an anti-Myc tag as detection antibody. The target protein was pulled down. Click image to enlarge

CPTC-PCNA-3 Immunoprecipitation

Result: Positive

Immuno-precipitation performed using CPTC-PCNA-3 as capture antibody against the over-expressed lysate of PCNA (Myc-tagged). Eluates were tested in automated western blot using the an anti-Myc tag as detection antibody. The target protein was pulled down.


Characterization SOP Files

  Immuno-MRM
Click to enlarge image Immuno-MRM chromatogram of CPTC-PCNA-3 antibody (see CPTAC assay portal for details: https://assays.cancer.gov/CPTAC-3243) Data provided by the Paulovich Lab, Fred Hutch (https://research.fredhutch.org/paulovich/en.html) Click image to enlarge

CPTC-PCNA-3 iMRM

Result: Positive

Immuno-MRM chromatogram of CPTC-PCNA-3 antibody (see CPTAC assay portal for details: https://assays.cancer.gov/CPTAC-3243)

Data provided by the Paulovich Lab, Fred Hutch (https://research.fredhutch.org/paulovich/en.html)


Characterization SOP Files

  NCI 60 Protein Array
Click to enlarge image Protein Array in which CPTC-PCNA-3 is screened against the NCI60 cell line panel for expression. Data is normalized to a mean signal of 1.0 and standard deviation of 0.5. Color conveys over-expression level (green), basal level (blue), under-expression level (red). Click image to enlarge

CPTC-PCNA-3 NCI 60 Protein Array

Result: Positive

Protein Array in which CPTC-PCNA-3 is screened against the NCI60 cell line panel for expression. Data is normalized to a mean signal of 1.0 and standard deviation of 0.5. Color conveys over-expression level (green), basal level (blue), under-expression level (red).


Characterization SOP Files

  Western Blot - Over-expressed transient protein in cell lysate
Click to enlarge image Western blot using CPTC-PCNA-3 as primary antibody against PCNA HEK293T cell transient overexpression lysate. The expected molecular weight is 28.8 kDa. Click image to enlarge

CPTC-PCNA-3 Western Blot (Over-Expressed Lysate)

Result: Positive

Western blot using CPTC-PCNA-3 as primary antibody against PCNA HEK293T cell transient overexpression lysate. The expected molecular weight is 28.8 kDa.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Western blot using CPTC-PCNA-3 as primary antibody against human PBMC (2), HeLa (3), Jurkat (4), A549 (5), MCF7 (6) and H226 (7) whole cell lysates. The expected molecular weight is 28.8 kDa. Cytochrome C was used as a loading control. Click image to enlarge

CPTC-PCNA-3 Western Blot (Cell Lysate)

Result: Positive

Western blot using CPTC-PCNA-3 as primary antibody against human PBMC (2), HeLa (3), Jurkat (4), A549 (5), MCF7 (6) and H226 (7) whole cell lysates. The expected molecular weight is 28.8 kDa. Cytochrome C was used as a loading control.


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Automated WB (Simple Western) using CPTC-PCNA-3 as primary antibody against the whole lysate of PBMC, HeLa, Jurkat, A549, MCF7 and H226. Expected MW is 65 KDa. The antibody recognizes the target in the tested lysates. The same cell lysates were probed with an anti-CytC antibody (bottom panel). Click image to enlarge

CPTC-PCNA-3 Simple Western (cell lysates)

Result: Positive

Automated WB (Simple Western) using CPTC-PCNA-3 as primary antibody against the whole lysate of PBMC, HeLa, Jurkat, A549, MCF7 and H226. Expected MW is 65 KDa. The antibody recognizes the target in the tested lysates. The same cell lysates were probed with an anti-CytC antibody (bottom panel).


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Automated WB (Simple Western) using CPTC-PCNA-3 as primary antibody against the whole lysate of breast, ovary, spleen, endometrium and lung tissues. Expected MW is 65 Kda. The antibody recognizes the target in the endometrium lysate and presuambly recognizes the target in the other tested lysates. The same cell lysates were probed with an anti-CytC antibody (bottom panel). Click image to enlarge

CPTC-PCNA-3 Simple Western (tissue lysates)

Result: Positive

Automated WB (Simple Western) using CPTC-PCNA-3 as primary antibody against the whole lysate of breast, ovary, spleen, endometrium and lung tissues. Expected MW is 65 Kda. The antibody recognizes the target in the endometrium lysate and presuambly recognizes the target in the other tested lysates. The same cell lysates were probed with an anti-CytC antibody (bottom panel).


Characterization SOP Files

  Western Blot - Tissue or Cell Lysate
Click to enlarge image Western blot using CPTC-PCNA-3 as primary antibody against human breast (2), ovary (3), spleen (4), endometrium (5), and lung (6) tissue lysates. The expected molecular weight is 28.8 kDa. Cytochrome C was used as a loading control. Click image to enlarge

CPTC-PCNA-3 Western Blot (Tissue Lysates)

Result: Presumed Positive (with additional bands)

Western blot using CPTC-PCNA-3 as primary antibody against human breast (2), ovary (3), spleen (4), endometrium (5), and lung (6) tissue lysates. The expected molecular weight is 28.8 kDa. Cytochrome C was used as a loading control.


Characterization SOP Files

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Antigen: Proliferating Cell Nuclear Antigen Peptide 3
Background

NCI Identification Number:

00190

Antigen Name:

Proliferating Cell Nuclear Antigen Peptide 3

CPTC Name:

CPTC-PCNA Peptide 3

Aliases:

DNA Polymerase Delta Auxiliary Protein; cyclin; Proliferating Cell Nuclear Antigen

Function:

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.

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. 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.

Chromosomal Localization:

20 pter-P12

Accession Number:

NP_002583.1

UniProt Accession Number:

P12004

DNA Source:

N/A

Immunogen:

Synthetic Peptide

Vector Name:

N/A

Extinction Coefficient:

Buffers:

Expressed Sequence:

AEDNADTLALVFEAPNQEK

Native Sequence:

Calculated Isoelectric Point:

0

Molecular Weight:

2090

Last Updated:

02/16/2017

Links

Characterization Data

SOPs

No SOPs available.

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