- NCBI Gene Id:
- Protein Name:
- Myeloid cell surface antigen CD33
- Alias Symbols:
- p67, SIGLEC3, FLJ00391, SIGLEC-3, CD33
- Replacement Item:
- This antibody may replace item sc-114798 from Santa Cruz Biotechnology.
- Description of Target:
- MOUSE ANTI HUMAN CD33
- Protein Size (# AA):
- IHC-AFF, FC, IP
- Tissue Tool:
- Find tissues and cell lines supported by DNA array analysis to express CD33.
- RNA Seq:
- Find tissues and cell lines supported by RNA-seq analysis to express CD33.
- The immunogen for anti-CD33 antibody: human AML cells
- Predicted Species Reactivity:
- Predicted Homology Based on Immunogen Sequence:
- Product Format:
- Purified IgG - liquid
- Reconstitution and Storage:
- Store at +4oC or at -20oC if preferred.
This product should be stored undiluted.
Storage in frost-free freezers is not recommended. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
- Printable datasheet for anti-CD33 antibody - OASA01906
- Application Info:
- Flow Cytometry: 1/10 - 1/50
Immunohistology - Frozen(1): 1/50 - 1/100
(1) See [*] for special conditions.
- Application Data:
- Application #1: Staining of human peripheral blood monocytes with Mouse anti Human CD33
Application #2: Staining of human peripheral blood monocytes with Mouse anti Human CD33:Alexa Fluor 488
Application #3: Staining of human peripheral blood monocytes with Mouse anti Human CD33:Alexa Fluor 647
Application #4: Staining of human peripheral blood monocytes with Mouse anti Human CD33:APC
Application #5: Staining of human peripheral blood monocytes with Mouse anti Human CD33:FITC
- Additional Information:
- Fusion Partners: Spleen cells from immunised BALB/c mice were fused with cells of the mouse NS1 myeloma cell line.
Histology Positive Control Tissue: Lymphatic tissue
- Preparation: Purified IgG prepared by affinity chromatography on Protein G from tissue culture supernatant
Preservative Stabilisers: 0.09% - Sodium Azide
- Approx Protein Conc: IgG concentration 1.0 mg/ml
Buffer Solutions: Phosphate buffered saline pH7.4
- Protocol Information:
- Citation: 1: Hernández-Caselles T, Martínez-Esparza M, Pérez-Oliva AB, Quintanilla-Cecconi AM, García-Alonso A, Alvarez-López DM, García-Peñarrubia P. A study of CD33 (SIGLEC-3) antigen expression and function on activated human T and NK cells: two isoforms of CD33 are generated by alternative splicing. J Leukoc Biol. 2006 Jan;79(1):46-58. PubMed PMID: 16380601.
Experiment Name: Immunoprecipitation (IP)
Experiment Background: IP assays using WM53 anti-CD33 mAb were performed to compare lymphoid and myeloid CD33 antigen
Experimental Steps: Cell-surface labeling and IPCell-surface proteins were labeled by biotinylation according to the protocol of the cellular labeling kit used (Boehringer Mannhein, Germany). Briefly, cells were washed twice in phosphate-buffered saline (PBS), pelleted, and resuspended in borate buffer at 107 cells/ml. Biotinylation reagent was added at 5 µg/ml and incubated under agitation at room temperature in the dark. After 15 min of incubation, NH4Cl (50 mM, final concentration) was added to stop the reaction and block free biotin. Then cells were washed twice in cold PBS and prepared for pervanadate stimulation or lysis and IP. Nonstimulated cells were resuspended in lysis buffer [50 mM Tris/ClH, pH 7.5, 150 mM NaCl, 1% Nonidet P-40 (NP-40), 1 mM phenylmethylsulfonyl fluoride (PMSF)] containing 2 ug/ml protease inhibitors (leupeptin, aprotinin, chemostatin, pepstatin). After 20 min on ice, lysates were centrifuged at 12,500 rpm for 15 min, precleared twice with protein A-Sepharose CL-4B beads (Amersham Pharmacia Biotech, Uppsala, Sweden), and immunoprecipitated overnight with protein A-Sepharose CL-4B beads, which had been preincubated previously with anti-CD33 mAb (Clone WM53). Subsequently, immunoprecipitates were washed five times with lysis buffer, resuspended in electrophoresis sample buffer, boiled at 95C for 5 min, and separated by SDS-polyacrylamide gel electrophoresis (PAGE). Finally, isolated immunoprecipitates were blotted onto polyvinylidene difluoride membranes, probed with horseradish peroxidase (HRP)-conjugated streptavidin, and developed using the enhanced chemiluminiscence (ECL) method (Amersham Pharmacia Biotech).
Other Reagents Used: Glutamax-I (L-alanyl-L-glutamine)
Number Of Protocols: 1
- Target Reference:
- 1. Favaloro, E.J. et al. (1987) Characterisation of monoclonal antibodies to the human myeloid-specific antigen 'gp67' (CD-33). Dis Markers. 5: 215-225.
2. Favaloro, E.J. et al. (1988) Further characterisation of myeloid antigens ('gp160,95', 'gp150' and 'gp67'): Investigation of epitopic heterogeneity and non-haemopoietic distribution using panels of monoclonal antibodies belonging to CD-11b, CD-13 and CD33. Br. J. Haematol. 69: 163-171.