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| Product Name | PCNA Recombinant Rabbit Monoclonal Antibody |
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| Antibody Type | Primary Antibodies |
| Product description | The proliferating cell nuclear antigen (PCNA), a protein synthesized in early G1 and S phases of the cell cycle, functions in cell cycle progression, DNA replication and DNA repair. In early S phase, PCNA exhibits granular distribution and is absent from the nucleoli; however, in late S phase, it relocates to the nucleoli. PCNA exists in two basic forms: one involved in ongoing DNA replication, which localizes specifically to the nucleus, and a second, soluble form, not implicated in constant synthesis. Interestingly, the latter form degrades in the presence of organic solvents, rendering it undetectable by histological methods in tissues using organic fixatives, and thus also providing a method of visualizing only the synthesizing form. |
| Immunogen | Synthetic peptide within Human PCNA aa 88-137 / 261. |
| Clonality | Monoclonal |
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| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:5000-1:20000 IHC:1:1000-1:10000 ICC/IF:1:2000 FC:1:200 | |
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/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:fb36g03 antibody |
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| Molecular Weight(MW) | 29kDa(Observed band size: 34kDa) |
| Cellular Localization | Nucleus. |
WB
Western blot analysis of PCNA on different lysates with Rabbit anti-PCNA antibody at 1/5,000 dilution. Lane 1: HEK-293 cell lysate, Lane 2: HeLa cell lysate, Lane 3: MCF7 cell lysate, Lane 4: NIH/3T3 cell lysate, Lane 5: C2C12 cell lysate, Lane 6: PC-12 cell lysate, Lysates/proteins at 15 µg/Lane. 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 at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit 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 Rabbit anti-PCNA antibody at 1/1,000 dilution. Lane 1: Mouse skin tissue lysate Lane 2: Mouse spleen tissue lysate Lysates/proteins at 40 µg/Lane. Exposure time: 10 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-Rabbit 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 tissue with Rabbit 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 small intestine tissue with Rabbit 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 human colon cancer tissue with Rabbit 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 human tonsil tissue with Rabbit 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.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling PCNA with Rabbit anti-PCNA antibody at 1/2,000 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 Rabbit anti-PCNA antibody at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit 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-Mouse IgG H&L (594) was 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 Rabbit IgG 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-Rabbit 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:1:5000-1:20000 IHC:1:1000-1:10000 ICC/IF:1:2000 FC:1:200 |
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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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