PCNA Recombinant Mouse Monoclonal Antibody [A6-G11-R]
cat.: HA601172
Product Type: Recombinant Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Cell, IP
Clonality: Monoclonal
Clone number: A6-G11-R
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/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: HeLa cell lysate, HEK-293 cell lysate, HCT 116 cell lysate, K-562 cell lysate, RAW264.7 cell lysate, C2C12 cell lysate, L6 cell lysate, mouse spleen tissue lysate, rat spleen tissue lysate, HeLa, Raji cell lysate, NIH/3T3 cell lysate, L-929 cell lysate, human liver tissue lysate, HepG2, human lymph node tissue, human melanoma tissue, rat spleen tissue.
Subcellular location: Nucleus
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
  IP
1:1,000-1:5,000
1:2,000
1:100-1:2,000
1-2μg/sample
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
Images
HA601172_1.jpg Fig1: Western blot analysis of PCNA on different lysates with Mouse anti-PCNA antibody (HA601172) at 1/20,000 dilution and competitor's antibody at 1/1,000 dilution.

Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: HEK-293 cell lysate (15 µg/Lane)
Lane 3: HCT 116 cell lysate (15 µg/Lane)
Lane 4: K-562 cell lysate (15 µg/Lane)
Lane 5: RAW264.7 cell lysate (15 µg/Lane)
Lane 6: C2C12 cell lysate (15 µg/Lane)
Lane 7: L6 cell lysate (15 µg/Lane)
Lane 8: Mouse spleen tissue lysate (15 µg/Lane)
Lane 9: Rat spleen tissue lysate (15 µg/Lane)

Predicted band size: 29 kDa
Observed band size: 34 kDa

Exposure time: 11 seconds; ECL: K1801;
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 (HA601172) at 1/20,000 dilution and competitor's antibody at 1/1,000 dilution were 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.
HA601172_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling PCNA with Mouse anti-PCNA antibody (HA601172) at 1/2,000 dilution and competitor's antibody at 1/1,600 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 (HA601172) at 1/2,000 dilution and competitor's antibody at 1/1,600 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.
HA601172_3.jpg Fig3: Western blot analysis of PCNA on different lysates with Mouse anti-PCNA antibody (HA601172) 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; ECL: K1801;

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 (HA601172) 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.
HA601172_4.jpg Fig4: Western blot analysis of PCNA on different lysates with Mouse anti-PCNA antibody (HA601172) 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; ECL: K1801;

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 (HA601172) 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.
HA601172_5.jpg Fig5: Immunocytochemistry analysis of HepG2 cells labeling PCNA with Mouse anti-PCNA antibody (HA601172) at 1/100 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 (HA601172) at 1/100 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.
HA601172_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human lymph node tissue with Mouse anti-PCNA antibody (HA601172) 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 (HA601172) 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.
HA601172_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human melanoma tissue with Mouse anti-PCNA antibody (HA601172) 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 (HA601172) 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.
HA601172_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Mouse anti-PCNA antibody (HA601172) 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 (HA601172) 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.
HA601172_9.jpg Fig9: Western blot analysis of PCNA on different lysates with Mouse anti-PCNA antibody (HA601172) 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: 10 seconds; ECL: K1801;

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 (HA601172) at 1/5,000 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.
HA601172_10.jpg Fig10: PCNA was immunoprecipitated from 0.2 mg HeLa cell lysate with HA601172 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA601172 at 1/10,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA601172 IP in HeLa cell lysate
Lane 3: Mouse IgG instead of HA601172 in HeLa cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 24 seconds; ECL: K1801
HA601172_11.jpg Fig11: PCNA was immunoprecipitated from 0.2 mg NIH/3T3 cell lysate with HA601172 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA601172 at 1/10,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: NIH/3T3 cell lysate (input)
Lane 2: HA601172 IP in NIH/3T3 cell lysate
Lane 3: Mouse IgG instead of HA601172 in NIH/3T3 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 5 seconds; ECL: K1802
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.