| Description | This product is a human monoclonal antibody that reacts with CD22. 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 | The extracellular domain of human CD22 and the full-length CD22 protein expressed on CHO cells |
| Epitope | Within the ECD domain. |
| 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 | ELISA; WB; DB; FC; FuncS |
| Application Notes | The antibody is recommended for detection of CD22 by FuncS assay. |
| ELISA | Enzyme-Linked Immunosorbent Assay Protocol |
| WB | Western Blot Protocol |
| FC | Flow Cytometry Protocol |
| Target | CD22 |
| Alternative Name | CD22; CD22 molecule; SIGLEC2; SIGLEC-2; B-cell receptor CD22; BL-CAM; CD22 antigen; T-cell surface antigen Leu-14; B-lymphocyte cell adhesion molecule; sialic acid binding Ig-like lectin 2; sialic acid-binding Ig-like lectin 2 |
| Gene ID | 933 |
| UniProt | P20273 |
| Related pathway | Hematopoietic Stem Cell Differentiation Pathways; Lineage-specific Markers; Innate Immune System |
| Research Area | Immunology; Cancer Research |
Figure 1 Anti-CD22 Monoclonal Antibody (V3S-0622-YC4301) in ELISA
ELISA analysis of V3S-0622-YC4301 was performed by coating with Human CD22 Protein (His Tag). Then blocked with BSA and incubated with V3S-0622-YC4301. 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.
Figure 1 Anti-CD22 Monoclonal Antibody (V3S-0622-YC4301) in ELISA
Figure 2 Anti-CD22 Monoclonal Antibody (V3S-0622-YC4301) in WB
Western blot analysis of V3S-0622-YC4301 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-0622-YC4301 (2 μg/mL) and HRP-conjugated goat anti-human IgG as a secondary antibody (1: 6000). Chemiluminescent detection was performed.
Figure 2 Anti-CD22 Monoclonal Antibody (V3S-0622-YC4301) in WB
Figure 3 Anti-CD22 Monoclonal Antibody (V3S-0622-YC4301) in DB
Dot Blot analysis of V3S-0622-YC4301 was performed by coating with Human CD22 Protein (His Tag).
V3S-0622-YC4301 incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-human IgG as a secondary antibody (1: 6000)
Figure 3 Anti-CD22 Monoclonal Antibody (V3S-0622-YC4301) in DB
Figure 4 Anti-CD22 Monoclonal Antibody (V3S-0622-YC4301) in SEC-HPLC
The purity of V3S-0622-YC4301 was greater than 99% 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-0622-YC4301) in SEC-HPLC
Figure 5 Anti-CD22 Monoclonal Antibody (V3S-0622-YC4301) in SDS-PAGE
SDS-PAGE analysis of V3S-0622-YC4301 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-0622-YC4301) in SDS-PAGE
Figure 6 Anti-CD22 Monoclonal Antibody (V3S-0622-YC4301) in WB
Western blot analysis of V3S-0622-YC4301 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-0622-YC4301) in WB
Figure 7 Anti-CD22 Monoclonal Antibody (V3S-0622-YC4301) in FACS
Flow cytometry analysis of Daudi cells (0.5x106) with purified antibody V3S-0622-YC4301 (5 μl). Cells were blocked with Human TruStain FcXTM and washed with 0.5% BSA-PBS. A FITC Goat anti-human IgG (5 μl) as a secondary antibody.
Figure 7 Anti-CD22 Monoclonal Antibody (V3S-0622-YC4301) in FACS
