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Description
Images & Validation
−| Tested Applications | ELISA, FC, IF, IHC-Fr, IHC-P, WB |
|---|---|
| Dilution Range | WB=1:500-2000, IHC-P=1:100-500, IF=1:100-500, Flow-Cyt=1ug/Test, IHC-F=1:100-500, ELISA=1:5000-10000 |
| Reactivity | Human, Mouse, Rat |
| Predicted Reactivity | Bovine, Gallus, Guinea pig, Porcine, Rabbit, Sheep |
Key Properties
−| Antibody Type | Primary Antibody |
|---|---|
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Immunogen | KLH conjugated synthetic peptide derived from human PCNA (151-261/261 aa) |
| Target | PCNA |
| Molecular Weight | 29 kDa |
| Purification | Affinity purified by Protein A |
| Conjugation | Unconjugated |
Storage & Handling
−| Storage | Shipped at 4°C. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles. |
|---|---|
| Form/Appearance | Liquid |
| Buffer/Preservatives | 0.01M TBS(pH7.4) with 1% rAlbumin, 0.03% Proclin300 and 50% Glycerol. |
| Concentration | 1mg/ml |
| Expiration Date | 12 months from date of receipt. |
| Disclaimer | For research use only |
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Blank control (blue line): U251 (blue). Primary Antibody (green line): Rabbit Anti-PCNA antibody (orb11248), Dilution: 3 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE, Dilution: 1 µg/Test. Protocol, The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2% BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Blank control: Jurkat. Primary Antibody (green line): Rabbit Anti-PCNA antibody (orb11248), Dilution: 1 ug/Test, Secondary Antibody: Goat anti-rabbit IgG-FITC, Dilution: 0.5 ug/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C. The cells were then incubated in 5% BSA to block non-specific protein-protein interactions for 30 min at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (Descending colon cancer), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (PCNA) Polyclonal Antibody, Unconjugated (orb11248) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human breast carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (PCNA) Polyclonal Antibody, Unconjugated (orb11248) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Incubation with (PCNA) Polyclonal Antibody, Unconjugated (orb11248) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Human esophageal cancer), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Incubation with (PCNA) Polyclonal Antibody, Unconjugated (orb11248) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (PCNA) Polyclonal Antibody, Unconjugated (orb11248) at 1:200 overnight at 4°C, followed by a conjugated secondary (orb868589) at [1:500] for 90 minutes and DAPI staining of the nuclei.

Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (PCNA) Polyclonal Antibody, Unconjugated at 1:200 overnight at 4°C, followed by a conjugated secondary (orb868589) at [1:500] for 90 minutes and DAPI staining of the nuclei.

Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (PCNA) Polyclonal Antibody, Unconjugated (orb11248) at 1:200 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human lung carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (PCNA) Polyclonal Antibody, Unconjugated (orb11248) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human rectal carcinoma), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (PCNA) Polyclonal Antibody, Unconjugated (orb11248) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse testis), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (PCNA) Polyclonal Antibody, Unconjugated (orb11248) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Lane 1: Hela (Human) Cell Lysate at 30 ug, Lane 2: MCF-7 (Human) Cell Lysate at 30 ug, Lane 3: MOLT-4 (Human) Cell Lysate at 30 ug, Lane 4: HepG2 (Human) Cell Lysate at 30 ug, Primary: Anti-PCNA (orb11248) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 32 kD, Observed band size: 32 kD.

Sample: Spleen (Mouse) Lysate at 40 ug, Primary: Anti-PCNA (orb11248) at 1/300 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 29 kD, Observed band size: 32 kD.

Tissue/Cell: human glioma tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-PCNA Polyclonal Antibody, Unconjugated (orb11248) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.
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Protocol Information
Zhao, Yong et al. Inhibition of peripubertal sheep mammary gland development by cysteamine through reducing progesterone and growth factor production Theriogenology, 89, 280-288 (2017)
PCNA Rabbit pAb (orb11248)
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