| Description | This product is a monoclonal antibody derived from Mouse (Mus musculus), which can specifically recognize Proliferating cell nuclear antigen. The antibody is expressed with mammalian cell transient expression system, serum-free and purified by affinity chromatography. The purity and integrity are tested via SDS-PAGE and SEC-HPLC analysis. Given an antigen, additional QC measures are also desired such as affinity testing and binding validation. Specifically, the antibody is provided in multiple formats for in vivo and in vitro assays. The Invivo version features greater than 95% purity, ultra-low endotoxin levels (<1 EU/mg or 0.1 EU/mg), and is preservative, stabilizer, and carrier protein-free. |
| Clonality | Monoclonal |
| Host Species | Mouse |
| Target Species | Human |
| Immunogen | Human His-PCNA |
| Affinity | KD = 35.4 nM, determined by surface plasmon resonance. |
| Isotype | IgG |
| Expression Species | HEK293F or CHO |
| Conjugation | None |
| Purity | >95%, determined by SDS-PAGE and/or SEC-HPLC |
| Endotoxin | <1 EU/mg, determined by LAL method |
| Purification | Protein A affinity purified |
| Sterility | 0.2 μM filtered |
| Formulation | PBS, pH 7.4 |
| Preservation | No preservatives |
| Stabilizer | No stabilizers |
| Storage | Store at 4⁰C within a week. For longer storage, aliquot and store at -20⁰C. |
| Application | DB; ELISA; FC; WB |
| Application Notes | The antibody is recommended for detection of PCNA by ELISA, FC, WB assays. |
| ELISA | Enzyme-Linked Immunosorbent Assay Protocol |
| WB | Western Blot Protocol |
| FC | Flow Cytometry Protocol |
Figure 1 Anti-PCNA Monoclonal Antibody (V3S-0522-YC1927) in WB
Western blot analysis of V3S-0522-YC1927 was performed by loading Human PCNA Protein (His Tag) in reduced condition Lane 1, Lane 2 and Lane 3 (0.1 μg, 0.3 μg and 0.6 μg respectively) onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least one hour. Membranes were probed with V3S-0522-YC1927 (2 μg/mL) and HRP-conjugated goat anti-mouse IgG as a secondary antibody (1:6000). Chemiluminescent detection was performed.
Figure 1 Anti-PCNA Monoclonal Antibody (V3S-0522-YC1927) in WB
Figure 2 Anti-PCNA Monoclonal Antibody (V3S-0522-YC1927) in DB
Dot Blot analysis of V3S-0522-YC1927 was performed by coating with Human PCNA Protein (His Tag).
V3S-0522-YC1927 incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-mouse IgG as a secondary antibody (1:6000)
Figure 2 Anti-PCNA Monoclonal Antibody (V3S-0522-YC1927) in DB
Figure 3 Anti-PCNA Monoclonal Antibody (V3S-0522-YC1927) in ELISA
ELISA analysis of V3S-0522-YC1927 was performed by coating with Human PCNA Protein (His Tag). Then blocked with BSA and incubated with V3S-0522-YC1927. The HRP-conjugated goat anti-mouse IgG as a secondary antibody (1:6000). Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.
Figure 3 Anti-PCNA Monoclonal Antibody (V3S-0522-YC1927) in ELISA
Figure 4 Anti-PCNA Monoclonal Antibody (V3S-0522-YC1927) in SDS-PAGE
SDS-PAGE analysis of V3S-0522-YC1927 in non-reduced (Lane 1) and β-mercaptoethanol-reduced (Lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result of different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively.
Figure 4 Anti-PCNA Monoclonal Antibody (V3S-0522-YC1927) in SDS-PAGE
Figure 5 Anti-PCNA Monoclonal Antibody (V3S-0522-YC1927) in SEC-HPLC
The purity of V3S-0522-YC1927 was greater than 90% as determined by SEC-HPLC.
Column: 3 µm, 7.8 x 300 nm
Mobile phase: 150 mM Sodium Phosphate Buffer, pH 7.0
Detection: UV 280 nm
Injection: 10 µl
Figure 5 Anti-PCNA Monoclonal Antibody (V3S-0522-YC1927) in SEC-HPLC
