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| Product Name | PCNA Mouse Monoclonal Antibody |
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| Antibody Type | Primary Antibodies |
| Product description | PCNA (Proliferating cell nuclear antigen) 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. It induces a robust stimulatory effect on the 3'-5' exonuclease and 3'-phosphodiesterase, but not apurinic-apyrimidinic (AP) endonuclease, APEX2 activities. |
| Immunogen | Recombinant protein within human PCNA aa 1-261. |
| Clonality | Monoclonal |
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| Isotype | IgG1 |
| Host Species | Mouse |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:1000-1:5000 IHC:1:2000 ICC/IF:1:500 FC:1:1000 | |
| Species Reactivity | HumanMouseRat |
| Concentration | 2mg/ml |
| Purification | Protein A |
| 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:fb36g05 antibody |
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| Molecular Weight(MW) | 29kD(Observed band size: 34kDa) |
| Cellular Localization | Nucleus |
| SwissProt ID | P12004 |
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WB
Western blot analysis of PCNA on different lysates with Mouse anti-PCNA antibody at 1/1,000 dilution. Lane 1: HCT 116 cell lysate (20 µg/Lane), Lane 2: HEK-293 cell lysate (20 µg/Lane), Lane 3: Raji cell lysate (20 µg/Lane), Lane 4: HeLa cell lysate (20 µg/Lane), Lane 5: K-562 cell lysate (20 µg/Lane), Lane 6: NIH/3T3 cell lysate (20 µg/Lane), Exposure time: 1 minutes 2 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/100,000 dilution was used for 1 hour at room temperature.
WB
Western blot analysis of PCNA on different lysates with Mouse anti-PCNA antibody at 1/1,000 dilution. Lane 1: NIH/3T3 cell lysate (20 µg/Lane), Lane 2: RAW264.7 cell lysate (20 µg/Lane), Lane 3: L-929 cell lysate (20 µg/Lane), Lane 4: C2C12 cell lysate (20 µg/Lane), Lane 5: Rat spleen tissue lysate (40 µg/Lane), Lane 6: Mouse spleen tissue lysate (40 µg/Lane), Lane 7: Human liver tissue lysate (40 µg/Lane), Exposure time: 7 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Mouse anti-PCNA antibody at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Mouse anti-PCNA antibody at 1/10,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/10,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Mouse anti-PCNA antibody at 1/10,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/10,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling PCNA with Mouse anti-PCNA antibody at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-PCNA antibody at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. beta Tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (594) were used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of HeLa cells labeling PCNA. Cells were fixed and permeabilized. Then stained with the primary antibody (1μg/mL) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:1000-1:5000 IHC:1:2000 ICC/IF:1:500 FC:1:1000 |
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| Form | Liquid |
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| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
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