CD33 Recombinant Rabbit Monoclonal Antibody [PSH02-51]
cat.: HA721827
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, FC
Clonality: Monoclonal
Clone number: PSH02-51
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: TF-1 cell lysate, TF-1, THP-1.
Subcellular location: Cell membrane; Peroxisome.
Recommended Dilutions:
  WB
  IF-Cell
  FC

1:2,000
1:500
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
HA721827_1.jpg Fig1: Western blot analysis of CD33 on different lysates with Rabbit anti-CD33 antibody (HA721827) at 1/2,000 dilution.

Lane 1: TF-1 cell lysate
Lane 2: Jurkat cell lysate (negative)
Lane 3: 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;

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 (HA721827) 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.
HA721827_2.jpg Fig2: Immunocytochemistry analysis of TF-1 (positive) and Jurkat (negative) labeling CD33 with Rabbit anti-CD33 antibody (HA721827) at 1/500 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 (HA721827) at 1/500 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.
HA721827_3.jpg Fig3: Immunocytochemistry analysis of THP-1 (positive) and Jurkat (negative) labeling CD33 with Rabbit anti-CD33 antibody (HA721827) at 1/500 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 (HA721827) at 1/500 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.
HA721827_4.jpg 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 (HA721827, 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”.