| Description | This product is a monoclonal antibody derived from Human (Homo sapiens), which can specifically recognize TNF receptor superfamily member 8. 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 | Human |
| Target Species | Human |
| Immunogen | Soluble CD30 |
| Affinity | KD = 1.2 nM, determined by surface plasmon resonance. |
| Isotype | IgG, κ |
| 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 | ELISA; WB; DB; FC; Cyt; FuncS |
| Application Notes | The antibody is recommended for detection of TNFRSF8 by FC, Cyt, FuncS assays. |
| ELISA | Enzyme-Linked Immunosorbent Assay Protocol |
| WB | Western Blot Protocol |
| FC | Flow Cytometry Protocol |
Figure 1 Anti-TNFRSF8 Monoclonal Antibody (V3S-0522-YC1963) in ELISA
ELISA analysis of V3S-0522-YC1963 was performed by coating with Human CD30/TNFRSF8 Protein (His Tag). Then blocked with BSA and incubated with V3S-0522-YC1963. The HRP-conjugated goat anti-human IgG as a secondary antibody (1: 8000). Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.
Lane 1: Reducing Antigen (0.1 μg)
Lane 2: Reducing Antigen (0.4 μg)
Lane 3: Reducing Antigen (0.6 μg)
Figure 1 Anti-TNFRSF8 Monoclonal Antibody (V3S-0522-YC1963) in ELISA
Figure 2 Anti-TNFRSF8 Monoclonal Antibody (V3S-0522-YC1963) in WB
Western blot analysis of V3S-0522-YC1963 was performed by loading Human CD30/TNFRSF8 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-YC1963 (2 μg/mL) and HRP-conjugated goat anti-human IgG as a secondary antibody (1: 6000). Chemiluminescent detection was performed.
Figure 2 Anti-TNFRSF8 Monoclonal Antibody (V3S-0522-YC1963) in WB
Figure 3 Anti-TNFRSF8 Monoclonal Antibody (V3S-0522-YC1963) in DB
Dot Blot analysis of V3S-0522-YC1963 was performed by coating with Human CD30/TNFRSF8 Protein (His Tag).
V3S-0522-YC1963 incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-mouse IgG as a secondary antibody (1: 6000)
NC: Negative control PC: Positive control
Figure 3 Anti-TNFRSF8 Monoclonal Antibody (V3S-0522-YC1963) in DB
Figure 4 Anti-TNFRSF8 Monoclonal Antibody (V3S-0522-YC1963) in SEC-HPLC
The purity of V3S-0522-YC1963 was greater than 95% 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 4 Anti-TNFRSF8 Monoclonal Antibody (V3S-0522-YC1963) in SEC-HPLC
Figure 5 Anti-TNFRSF8 Monoclonal Antibody (V3S-0522-YC1963) in SDS-PAGE
SDS-PAGE analysis of V3S-0522-YC1963 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 5 Anti-TNFRSF8 Monoclonal Antibody (V3S-0522-YC1963) in SDS-PAGE
