RB780 Biosimilar Anti-Human CD20

BD Horizon™ RB780 Biosimilar Anti-Human CD20

Name: RB780 Biosimilar Anti-Human CD20
Brand: RB780 Biosimilar Anti-Human CD20
SKU: 570975

Clone Obinutuzumab297.rMAb (also known as Obinutuzumab N297A Biosimilar.rMAb) (RUO)

Flow cytometric analysis of Human CD20 (Obinutuzumab Biosimilar) expression on Human peripheral blood leukocytes. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with either BD OptiBuild™ RB780 Human IgG1 (N297A), κ Isotype Control (Cat. No. 756229; Left Plot) or BD Horizon™ RB780 Biosimilar Anti-Human CD20 (Obinutuzumab) antibody (Cat. No. 570975/571046; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD20 (Obinutuzumab Biosimilar) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocyte populations. Flow cytometric analysis was performed using a BD FACSymphony™ A5 Cell Analyzer System and FlowJo™ Software.

Flow cytometric analysis of Human CD20 (Obinutuzumab Biosimilar) expression on Human peripheral blood lymphocytes. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219) and stained with BD Horizon™ BV421 Mouse Anti-Human CD19 antibody (Cat. No. 562440) and with either BD OptiBuild™ RB780 Human IgG1 (N297A), κ Isotype Control (Cat. No. 756229; Left Plot) or BD Horizon™ RB780 Biosimilar Anti-Human CD20 (Obinutuzumab) antibody (Cat. No. 570975/571046; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD20 (Obinutuzumab Biosimilar) [or Ig Isotype control staining] versus CD19 was derived from gated events with the forward and side light-scatter characteristics of viable lymphocytes. Flow cytometric analysis was performed using a BD FACSymphony™ A5 Cell Analyzer System and FlowJo™ Software.

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Product Details

Brand: BD Horizon™

Alternative Name: B1; Bp35; LEU-16; MS4A1

Reactivity: Human (QC Testing)

Isotype: Human IgG1, κ

Immunogen: Human CD20

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl/test

Entrez Gene ID: 931

RRID: AB_3686155

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

Regulatory Status: RUO

Preparation And Storage

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 unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

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.

Product Notices

When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
An isotype control should be used at the same concentration as the antibody of interest.
Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
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.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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.
Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
For U.S. patents that may apply, see bd.com/patents.

Companion Products

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

RB780 Human IgG1 (N297A), κ Isotype Control RUO

Size 50 µg Cat No. 756229

570975 Rev. 1

Antibody Details

Obinutuzumab297.rMAb

The Obinutuzumab297.rMAb (also known as, Obinutuzumab N297A Biosimilar.rMAb) is a research grade humanized recombinant human IgG1, kappa antibody that specifically recognizes the extracellular domain of human CD20 similarly to the therapeutic Obinutuzumab antibody. The Obinutuzumab297.rMAb uses the same variable region sequences as Obinutuzumab, combined with constant region sequences derived from consensus sequences of human IgG1, kappa. The asparagine at position 297 of the constant heavy chain has been replaced with alanine (N297A) to further reduce Fc receptor interactions of this reagent. Obinutuzumab triggers target cell death through multiple mechanisms including complement-dependent cytotoxicity (CDC), antibody-dependent cellular cytotoxicity (ADCC), and apoptosis. The therapeutic Obinutuzumab has been approved to treat human B cell malignancies, autoimmune disorders, and transplant rejection. CD20 is a 33-37 kDa, unglycosylated four-transmembrane phosphoprotein with a cytoplasmic N-terminus and C-terminus that is encoded by MS4A1 (Membrane-spanning 4-domains, subfamily A, member 1). CD20 is expressed on pre-B-cells, naive and activated B cells, memory B cells, and at lower levels on a small subset of T cells but is absent from plasmablasts and plasma cells. It is also expressed on most malignant B cells and CD20-positive T-cell lymphomas. CD20 functions in mediating calcium transport and B cell activation, differentiation, and survival.

The Obinutuzumab297.rMAb is intended for research use only. It is not intended for use in therapeutic or diagnostic procedures for humans or animals.

570975 Rev. 1

Format Details

RB780

The BD Horizon RealBlue™ 780 (RB780) Dye is part of the BD family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 781-nm. Driven by BD innovation, RB780 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.

Format: RB780

Excitation Source: Blue 488 nm

Excitation Max: 498 nm

Emission Max: 781 nm

570975 Rev.1

Citations & References

View product citations for antibody "570975" on CiteAb

Development References (5)

Almasri NM, Duque RE, Iturraspe J, Everett E, Braylan RC. Reduced expression of CD20 antigen as a characteristic marker for chronic lymphocytic leukemia. Am J Hematol. 1992; 40:259-263. (Biology).
Brekke OH, Sandlie I. Therapeutic antibodies for human diseases at the dawn of the twenty-first century.. Nat Rev Drug Discov. 2003; 2(1):52-62. (Biology). View Reference
Hultin LE, Hausner MA, Hultin PM, Giorgi JV. CD20 (pan-B cell) antigen is expressed at a low level on asubpopulation of human T lymphocytes. Cytometry. 1993; 14:196-204. (Biology).
Tobinai K, Klein C, Oya N, Fingerle-Rowson G. A Review of Obinutuzumab (GA101), a Novel Type II Anti-CD20 Monoclonal Antibody, for the Treatment of Patients with B-Cell Malignancies.. Adv Ther. 2017; 34(2):324-356. (Biology). View Reference
Zola H, Swart B, Nicholson I, Voss E. CD20. In: Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007:72.

View All (5)

570975 Rev. 1

Please refer to Support Documents for Quality Certificates

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.