PCNA Recombinant antibody

PCNA Uni-rAb^TM Recombinant Antibody for WB, IHC, IF-P, FC (Intra), ELISA

Host / Isotype

Rabbit / IgG

Reactivity

human, mouse, rat

Applications

WB, IHC, IF-P, FC (Intra), ELISA

Conjugate

Unconjugated

CloneNo.

240306B9

Cat no : 81302-6-RR

Synonyms

proliferating cell nuclear antigen, Cyclin, 240306B9

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Validation Data Gallery

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  WB analysis using 81302-6-RR

  Various lysates were subjected to SDS PAGE followed by western blot with 81302-6-RR (PCNA antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours.

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  IHC staining of human stomach cancer using 81302-6-RR

  Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 81302-6-RR (PCNA antibody) at dilution of 1:4000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IHC staining of human stomach cancer using 81302-6-RR

  Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 81302-6-RR (PCNA antibody) at dilution of 1:4000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  IF Staining of mouse testis using 81302-6-RR

  Immunofluorescent analysis of (4% PFA) fixed paraffin-embedded mouse testis tissue using PCNA antibody (81302-6-RR, Clone: 240306B9 ) at dilution of 1:500 and Multi-rAb CoraLite ® Plus 488-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L) (RGAR002). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

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  FC experiment of HeLa using 81302-6-RR

  1x10^6 HeLa cells were intracellularly stained with 0.25 ug PCNA Recombinant antibody (81302-6-RR, Clone:240306B9) and APC-Conjugated AffiniPure Goat Anti-Rabbit IgG(H+L)(red), or 0.25 ug Isotype Control (blue). Cells were fixed and permeabilized with Transcription Factor Staining Buffer Kit (PF00011).

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Tested Applications

• Positive WB detected in: HeLa cells, HEK-293 cells, Jurkat cells, A431 cells, MCF-7 cells, NIH/3T3 cells, HSC-T6 cells
• Positive IHC detected in: human stomach cancer tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: mouse testis tissue
• Positive FC (Intra) detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunohistochemistry (IHC): IHC : 1:2000-1:8000
• Immunofluorescence (IF)-P: IF-P : 1:250-1:1000
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

The immunogen of 81302-6-RR is PCNA Fusion Protein expressed in E. coli.

• Tested Reactivity: human, mouse, rat
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen: PCNA fusion protein Ag0277
• Full Name: proliferating cell nuclear antigen
• Calculated Molecular Weight: 29 kDa/31 kDa
• Observed Molecular Weight: 36 kDa
• GenBank Accession Number: BC000491
• Gene Symbol: PCNA
• Gene ID (NCBI): 5111
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Proliferating Cell Nuclear Antigen, commonly known as PCNA, is a protein that acts as a processivity factor for DNA polymerase δ in eukaryotic cells. 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. PCNA induces a robust stimulatory effect on the 3'-5' exonuclease and 3'-phosphodiesterase, but not apurinic-apyrimidinic (AP) endonuclease, APEX2 activities. It has to be loaded onto DNA in order to be able to stimulate APEX2. PCNA protein is highly conserved during evolution; the deduced amino acid sequences of rat and human differ by only 4 of 261 amino acids. PCNA has been used as loading control for proliferating cells. The calculated molecular weight of PCNA is 29 kDa, but modified PCNA is 36kDa (PMID: 1358458).