BUV395 Mouse Anti-Human CD33

This product is the replacement for 740293.

BD Horizon™ BUV395 Mouse Anti-Human CD33

Name: BUV395 Mouse Anti-Human CD33
Brand: BD Horizon™
SKU: 568374

Clone WM53 (also known as WM-53)

(RUO)

Multiparameter flow cytometric analysis of CD33 expression on Human peripheral blood leucocyte populations. Whole blood was stained with either BD Horizon™ BUV395 Mouse IgG1, k Isotype Control (Cat. No. 563547; Left Plot) or BD Horizon™ BUV395 Mouse Anti-Human CD33 antibody (Cat. No. 568374; Right Plot) at 0.25 µg/test. The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The bivariate pseudocolor density plot showing the correlated expression of CD33 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the side and forward light-scattering characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

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Product Details

Brand: BD Horizon™

Alternative Name: Siglec-3; SIGLEC3; Sialic acid-binding Ig-like lectin 3; p67; gp67; My9

Reactivity: Human (QC Testing)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: Acute Myeloid Leukemia Blasts

Application: Flow cytometry (Routinely Tested)

Concentration: 0.2 mg/ml

Workshop Number: IV M505

Entrez Gene ID: 945

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: RUO

Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Product Notices

Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
Since applications vary, each investigator should titrate the reagent to obtain optimal results.
An isotype control should be used at the same concentration as the antibody of interest.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 8,575,303; 8,354,239.
BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Brilliant Stain Buffer Plus RUO

Size 1000 Tests Cat No. 566385

Brilliant Stain Buffer RUO

Size 1000 Tests Cat No. 566349

BUV395 Mouse IgG1, k Isotype Control RUO

Size 50 µg Cat No. 563547

Lysing Buffer RUO

Size 100 mL Cat No. 555899

View Details

568374 Rev. 1

Antibody Details

WM53

The WM53 monoclonal antibody specifically recognizes CD33 which is also known as Sialic acid-binding Ig-like lectin 3 (Siglec-3) or gp67. CD33 is a 67 kDa type I transmembrane glycoprotein that is variably expressed on myeloid progenitors, monocytes, macrophages, dendritic cells, neutrophils, basophils, mast cells, and on some activated T cells and NK cells. Normal lymphocytes, platelets, erythrocytes and pluripotent hematopoietic stem cells do not express the CD33 antigen. This glycoprotein reportedly functions as a sialic acid-dependent cell adhesion molecule and this function can be modulated by endogenous sialoglycoconjugates when CD33 is expressed on the membrane.

568374 Rev. 1

Format Details

BUV395

The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BUV395

Excitation Source: Ultraviolet 355 nm

Excitation Max: 348 nm

Emission Max: 395 nm

568374 Rev.1

Citations & References

Development References (7)

Favaloro EJ, Bradstock KF, Kabral A, Grimsley P, Berndt MC. Characterization of monoclonal antibodies to the human myeloid-differentiation antigen, 'gp67' (CD-33). Dis Markers. 1987; 5(4):215-225. (Immunogen: Blocking, Flow cytometry, Fluorescence microscopy, Immunoprecipitation, Radioimmunoassay). View Reference
Favaloro EJ, Bradstock KF, Kabral A, Grimsley P, Zowtyj H, Zola H. Further characterization of human myeloid antigens (gp160,95; gp150; gp67): investigation of epitopic heterogeneity and non-haemopoietic distribution using panels of monoclonal antibodies belonging to CD-11b, CD-13 and CD-33. Br J Haematol. 1988; 69(2):163-171. (Clone-specific: Blocking, Flow cytometry, Immunofluorescence, Immunohistochemistry, Radioimmunoassay). View Reference
Favaloro EJ, Moraitis N, Koutts J, Exner T, Bradstock KF. Endothelial cells and normal circulating haemopoietic cells share a number of surface antigens. Thromb Haemost. 1989; 61(2):217-224. (Biology). View Reference
Freeman SD, Kelm S, Barber EK, Crocker PR. Characterization of CD33 as a new member of the sialoadhesin family of cellular interaction molecules. Blood. 1995; 85(8):2005-2012. (Biology). View Reference
Hernández-Caselles T, Martínez-Esparza M, Pérez-Oliva AB, et al. 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; 79(1):46-58. (Clone-specific: Flow cytometry). View Reference
Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:1-1182.
Nakamura Y, Noma M, Kidokoro M, et al. Expression of CD33 antigen on normal human activated T lymphocytes.. Blood. 1994; 83(5):1442-3. (Biology). View Reference

View All (7)

568374 Rev. 1

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Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.