CD33 Recombinant Rabbit Monoclonal Antibody [PSH02-50]
cat.: HA721826
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human |
| Applications: | WB, IF-Cell, FC |
| Clonality: | Monoclonal |
| Clone number: | PSH02-50 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 40 kDa |
| Isotype: | IgG |
| Immunogen: | Recombinant protein within human CD33 aa 1-282 / 364 (P20138). |
| Positive control: | THP-1 cell lysate, TF-1 cell lysate, HL-60 cell lysate, TF-1. |
| Subcellular location: | Cell membrane; Peroxisome. |
| Recommended Dilutions:
WB IF-Cell FC |
1:2,000 1:200 1:1,000 |
| Uniprot #: | SwissProt: P20138 Human |
| Alternative names: | CD 33 CD33 CD33 antigen (gp67) CD33 antigen CD33 molecule CD33_HUMAN FLJ00391 gp67 My9 Myeloid cell surface antigen CD33 Myeloid cell surface antigen CD33 precursor Myeloid differentiation antigen CD33 p67 Sialic acid binding Ig like lectin 3 Sialic acid binding immunoglobulin like lectin 3 Sialic acid-binding Ig-like lectin 3 SIGLEC 3 Siglec-3 SIGLEC3 |
Images
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Fig1:
Western blot analysis of CD33 on different lysates with Rabbit anti-CD33 antibody (HA721826) at 1/2,000 dilution. Lane 1: THP-1 cell lysate Lane 2: TF-1 cell lysate Lane 3: HL-60 cell lysate Lane 4: Jurkat cell lysate (negative) Lane 5: SK-MEL-28 cell lysate (negative) Lysates/proteins at 20 µg/Lane. Predicted band size: 40 kDa Observed band size: 70 kDa Exposure time: 3 minutes; ECL: K1802; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721826) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig2:
Western blot analysis of CD33 on different lysates with Rabbit anti-CD33 antibody (HA721826) at 1/1,000 dilution. Lane 1: THP-1 cell lysate Lane 2: THP-1 cell lysate treated with deglycosylation Predicted band size: 40 kDa Observed band size: 40/70 kDa (Glycosylated) Exposure time: 3 minutes; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721826) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
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Fig3:
Immunocytochemistry analysis of TF-1 (positive) and Jurkat (negative) labeling CD33 with Rabbit anti-CD33 antibody (HA721826) at 1/200 dilution. Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD33 antibody (HA721826) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
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Fig4:
Flow cytometric analysis of TF-1 cells labeling CD33. Cells were washed twice with cold PBS and resuspend. Then stained with the primary antibody (HA721826, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.