| Description | This product is a monoclonal antibody derived from Human (Homo sapiens), which can specifically recognize Fms Related Tyrosine Kinase 3. 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 |
| Epitope | The epitope is domain 5 of FLT3. |
| Isotype | IgG1, 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 | WB; ELISA; Neut |
| Application Notes | The antibody is recommended for detection of FLT3 by ELISA, Neut assays. |
| ELISA | Enzyme-Linked Immunosorbent Assay Protocol |
| WB | Western Blot Protocol |
| FC | Flow Cytometry Protocol |
Figure 1 Anti-FLT3 Monoclonal Antibody (V3S-0522-YC556) in SDS-PAGE
SDS-PAGE analysis of V3S-0522-YC556 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. And non-reduced protein migrates as about 180 kDa.
Figure 1 Anti-FLT3 Monoclonal Antibody (V3S-0522-YC556) in SDS-PAGE
Figure 2 Anti-FLT3 Monoclonal Antibody (V3S-0522-YC556) in WB
Western blot analysis of V3S-0522-YC556 was performed by loading 2 µg onto a 12% Tris-HCl polyacrylamide gel in non-reduced (lane 1) and β-mercaptoethanol-reduced (lane 2) conditions. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least one hour. Membranes were probed with Goat Anti-Human IgG HRP secondary antibody at a dilution of 1:8,000 for 75 minutes. Chemiluminescent detection was performed.
Figure 2 Anti-FLT3 Monoclonal Antibody (V3S-0522-YC556) in WB
Figure 3 Anti-FLT3 Monoclonal Antibody (V3S-0522-YC556) in SEC-HPLC
The purity of V3S-0522-YC556 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 7.3
Detection: UV 280 nm
Injection: 25 µl
Figure 3 Anti-FLT3 Monoclonal Antibody (V3S-0522-YC556) in SEC-HPLC
Figure 4 Anti-FLT3 Monoclonal Antibody (V3S-0522-YC556) in WB
Western blot analysis was performed by loading 1 µg (lane 1) and 1 µg (lane 2) recombinant human FLT3 antigen onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least 1 hour. Membranes were probed with TLT3 antibody (V3S-0522-YC556) at a dilution of 1:1000 overnight at 4°C on a rocking platform. Then probed with Goat anti-Human IgG HRP secondary antibody at a dilution of 1:8,000 for one hour. Chemiluminescent was detected.
Figure 4 Anti-FLT3 Monoclonal Antibody (V3S-0522-YC556) in WB
Figure 5 Anti-FLT3 Monoclonal Antibody (V3S-0522-YC556) in ELISA
ELISA analysis of V3S-0522-YC556 was performed by coating with recombinant human FLT3 antigen. Then blocked with BSA, and incubated with TLT3 antibody (V3S-0522-YC556) at a starting concentration of 1µg/mL. The HRP-conjugated goat anti-human IgG as a secondary antibody. Detection was performed using TMB substrate and stopped with sulfuric acid. The absorbances were read on a spectrophotometer at 450 nm.
Figure 5 Anti-FLT3 Monoclonal Antibody (V3S-0522-YC556) in ELISA
