| Description | This product is a recombinant Mouse antibody provided by Creative Biolabs. The antibody is specific for CD22. It can be used for CD22 detection in Enzyme-Linked Immunosorbent Assay and Inhibition Assay. It was expressed in mammalian cells (293F or CHO) with antibody encoding genes and purified by affinity chromatography. Each lot of this antibody is quality control tested by SDS-PAGE and SEC-HPLC analysis. For highly sensitive assays, we recommend the ultra purified form of the product, which has a lower endotoxin limit than standard antibody, less than 1 EU/mg or even 0.1 EU/mg. |
| Clonality | Monoclonal |
| Host Species | Mouse |
| Target Species | Human |
| Isotype | IgG |
| Expression Species | HEK293F or CHO cell line |
| Conjugation | Unconjugated, also available for Biotin, HRP, FITC and PE-labeled form. |
| Purity | >95%, determined by SDS-PAGE and SEC-HPLC |
| Endotoxin | <1 EU/mg, determined by LAL method |
| Purification | Purified with Protein A or G affinity chromatography |
| Sterility | 0.2 μM filtered |
| Formulation | PBS, pH 7.4 |
| Preservation | No preservatives |
| Stabilizer | No stabilizers |
| Storage | Store at 4°C within one or two weeks. Store at -20°C for long term. Avoid repeated freeze/thaw cycles. Refer to the COA file for specifics. |
| Application | WB; DB; FC; ELISA; Inhib |
| ELISA | Enzyme-Linked Immunosorbent Assay Protocol |
| WB | Western Blot Protocol |
| FC | Flow Cytometry Protocol |
Figure 1 Anti-CD22 Monoclonal Antibody (V3S-1222-YC1463) in ELISA
ELISA analysis of V3S-1222-YC1463 was performed by coating with Human CD22 Protein (His Tag). Then blocked with BSA and incubated with V3S-1222-YC1463. The HRP-conjugated goat anti-mouse 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.
Figure 1 Anti-CD22 Monoclonal Antibody (V3S-1222-YC1463) in ELISA
Figure 2 Anti-CD22 Monoclonal Antibody (V3S-1222-YC1463) in WB
Western blot analysis of V3S-1222-YC1463 was performed by loading Human CD22 Protein (His Tag) in reduced condition Lane 1 (0.8 μg) 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-1222-YC1463 (2 μg/mL) and HRP-conjugated goat anti-mouse IgG as a secondary antibody (1: 6000). Chemiluminescent detection was performed.
Figure 2 Anti-CD22 Monoclonal Antibody (V3S-1222-YC1463) in WB
Figure 3 Anti-CD22 Monoclonal Antibody (V3S-1222-YC1463) in DB
Dot Blot analysis of V3S-1222-YC1463 was performed by coating with Human CD22 Protein (His Tag).
V3S-1222-YC1463 incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-mouse IgG as a secondary antibody (1: 6000)
Figure 3 Anti-CD22 Monoclonal Antibody (V3S-1222-YC1463) in DB
Figure 4 Anti-CD22 Monoclonal Antibody (V3S-1222-YC1463) in SEC-HPLC
The purity of V3S-1222-YC1463 was greater than 96% 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 4 Anti-CD22 Monoclonal Antibody (V3S-1222-YC1463) in SEC-HPLC
Figure 5 Anti-CD22 Monoclonal Antibody (V3S-1222-YC1463) in SDS-PAGE
SDS-PAGE analysis of V3S-1222-YC1463 in non-reduced and β-mercaptoethanol-reduced 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 175-270 kDa.
Figure 5 Anti-CD22 Monoclonal Antibody (V3S-1222-YC1463) in SDS-PAGE
Figure 6 Anti-CD22 Monoclonal Antibody (V3S-1222-YC1463) in WB
Western blot analysis of V3S-1222-YC1463 was performed by loading 2 µg onto a 12% Tris-HCl polyacrylamide gel in reduced (Lane 2) and non-reduced (Lane 1) conditions.
Figure 6 Anti-CD22 Monoclonal Antibody (V3S-1222-YC1463) in WB
Figure 7 Anti-CD22 Monoclonal Antibody (V3S-1222-YC1463) in FACS
Flow cytometry analysis of Daudi cells (0.5x106) with purified antibody V3S-1222-YC1463 (5 μl). Cells were blocked with Human TruStain FcXTM and washed with 0.5% BSA-PBS. An Alexa Fluor 488 goat anti-mouse IgG(H+L) (5 μl) as a secondary antibody.
Figure 7 Anti-CD22 Monoclonal Antibody (V3S-1222-YC1463) in FACS
