Product Type: | Mouse monoclonal IgG1, primary antibodies |
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Species reactivity: | Human, Mouse, Rat |
Applications: | WB, IHC-P, FC, IF-Cell |
Clonality: | Monoclonal |
Clone number: | A6-G11 |
Form: | Liquid |
Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
Storage buffer: | 1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
Concentration: | 2ug/ul |
Purification: | Protein A affinity purified. |
Molecular weight: | Predicted band size: 29 kDa |
Isotype: | IgG1 |
Immunogen: | Recombinant protein within human PCNA aa 1-261. |
Positive control: | HCT 116 cell lysate, HEK-293 cell lysate, Raji cell lysate, HeLa cell lysate, K-562 cell lysate, NIH/3T3 cell lysate, RAW264.7 cell lysate, L-929 cell lysate, C2C12 cell lysate, rat spleen tissue lysate, mouse spleen tissue lysate, human liver tissue lysate, HeLa, human colon cancer tissue, mouse testis tissue, rat testis tissue. |
Subcellular location: | Nucleus |
Recommended Dilutions:
WB IHC-P FC IF-Cell |
1:1,000-1:5,000 1:10,000 1:1,000 1:500 |
Uniprot #: | SwissProt: P12004 Human | P17918 Mouse | P04961 Rat |
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:fb36g05 |
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Fig1:
Western blot analysis of PCNA on different lysates with Mouse anti-PCNA antibody (EM111201) 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) Predicted band size: 29 kDa Observed band size: 34 kDa 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 (EM111201) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/100,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Western blot analysis of PCNA on different lysates with Mouse anti-PCNA antibody (EM111201) 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) Predicted band size: 29 kDa Observed band size: 34 kDa 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 (EM111201) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig3:
Western blot analysis of PCNA on different lysates with Rabbit anti-PCNA antibody (EM111201) at 1/5,000 dilution. Lane 1: HeLa-si NT cell lysate Lane 2: HeLa-si PCNA cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 29 kDa Observed band size: 34 kDa Exposure time: 24 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 (EM111201) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig4:
Immunocytochemistry analysis of HeLa cells labeling PCNA with Mouse anti-PCNA antibody (EM111201) 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 (EM111201) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution. |
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Fig5:
Flow cytometric analysis of HeLa cells labeling PCNA. Cells were fixed and permeabilized. Then stained with the primary antibody (EM111201, 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 (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
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Fig6:
Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-PCNA antibody (EM111201) 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 (EM111201) 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. |
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Fig7:
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-PCNA antibody (EM111201) 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 (EM111201) 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. |
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Fig8:
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-PCNA antibody (EM111201) 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 (EM111201) 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. |