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Spark Blue™ 574 anti-human CD20 (Flexi-Fluor™) Antibody

Pricing & Availability
Clone
2H7 (See other available formats)
Regulatory Status
RUO
Workshop
IV B201
Other Names
B1, Bp35
Isotype
Mouse IgG2b, κ
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Product Citations
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Compare all formats See Spark Blue™ 574 spectral data
Cat # Size Price Quantity Save
285019 50 µg 394 CHF
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Description

CD20 is a 33-37 kD, four transmembrane spanning protein, also known as B1 and Bp35. CD20 is expressed on pre-B-cells, resting and activated B cells (not plasma cells), some follicular dendritic cells, and at low levels on a T cell subset. CD20 is heavily phosphorylated on activated B cells and malignant B cells. Homo-oligomeric complexes of CD20 are thought to form Ca2+ conductive ion channels in the plasma membrane of B cells. The CD20 molecule is involved in B-cell activation and is associated with various Src family kinases (Lyn, Lck, Fyn). It exists in a complex with MHC class I and II, CD53, CD81, and CD82.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Cynomolgus, Rhesus
Reported Reactivity
Baboon, Capuchin Monkey, Chimpanzee, Pigtailed Macaque, Squirrel Monkey
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Human tonsillar B cells
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography and conjugated with Spark Blue™ 574 under optimal conditions.
Concentration
0.2 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

FC
Recommended Usage
Flexi-Fluors™ are provided at a standard 0.2 mg/mL concentration. We recommend titrating this reagent to determine the optimal concentration for each application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL. We recommend testing a range of concentrations starting from 10 µg/mL.

For example, make five 1:1 serial dilutions of the 0.2 mg/mL antibody. Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/mL) to test six concentrations -- 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration.
Excitation Laser
Blue Laser (488 nm)
Application Notes

The epitope recognized by clone 2H7 has been mapped to the sequence YNCEPANPSEKNSPST which lies in the large extracellular loop of human CD20. Additional reported applications (for the relevant formats) include: immunoprecipitation4 and immunohistochemical staining of acetone-fixed frozen sections5.

Additional Product Notes

For more information about Flexi-Fluors™, visit our Flexi-Fluor™ page and review FAQs associated with this product line.

Application References

(PubMed link indicates BioLegend citation)
  1. Schlossman S, et al. 1995. Leucocyte Typing V. Oxford University Press. New York.
  2. Knapp W, et al. 1989. Leucocyte Typing IV. Oxford University Press. New York.
  3. McMichael A, et al. Eds. 1987. Leucocyte Typing III Oxford University Press. New York.
  4. Polyak MJ, et al. 2002. Blood 99:3256. (IP)
  5. Mack CL, et al. 2004. Pediatr. Res. 56:79. (IHC)
RRID
AB_3106284 (BioLegend Cat. No. 285019)

Antigen Details

Structure
Four transmembrane protein (TM4SF), heavily phosphorylated after activation, 33-37 kD
Distribution

B cell, T cell subsets

Function
B cell activation
Ligand/Receptor
Src family tyrosine kinases, MHC class I, II, CD53, CD81, CD82
Cell Type
B cells, T cells
Biology Area
Costimulatory Molecules, Immunology
Molecular Family
CD Molecules
Antigen References

1. Hultin L, et al. 1993. Cytometry 14:196.
2. Tedder T, et al. 1994. Immunol. Today 15:450.

Gene ID
931 View all products for this Gene ID
UniProt
View information about CD20 on UniProt.org

Related FAQs

What is the difference between two anti human CD20 clones 2H7 and 1412?

For clone 1412: This clone specifically recognizes cytoplasmic domain of CD20 and thus can only be used for intracellular flow cytometry. In this instance you will need to include the fixation and permeabilization steps. Please follow the intracellular flow cytometry staining protocol.

For clone 2H7: This clone is ok for regular surface staining for CD20 and there is no need for any fixation and permeabilization steps.

Our technical protocols can be found here.

What are Flexi-Fluors?

Flexi-Fluors are rapidly made-to-order conjugated antibodies. The technology, manufacturing processes, and specifications used to create Flexi-Fluors are the same as our regular catalog products. However, the optimal concentration and performance of each Flexi-Fluor must be determined by the customer. 

How quickly will I receive my order?

We aim to ship Flexi-Fluors within 2-3 weeks of receipt of your order. However, depending on your location, shipping times may vary. 

How are Flexi-Fluors different from regular catalog products?

Flexi-Fluors are made on demand, specifically for you. Flexi-Fluors are manufactured using the same high-quality standards, and specifications as other catalog products. For faster delivery, Flexi-Fluors are not tested by flow cytometry to determine optimal concentrations or evaluate performance. This testing needs to be performed by the customer.

How do I determine the optimal concentration for using my Flexi-Fluor? How should I titrate my antibody?

Flexi-Fluors are provided at a standard 0.2 mg/mL concentration. We recommend that you titrate your antibody to determine the optimal concentration to use for your application. For many flow cytometry applications, conjugated antibodies perform well at concentrations ranging from 0.03 to 1.0 µg per million cells in 100 µL volume. We recommend that you test a range of concentrations starting from 10 µg/mL. 

 

For example, make five 1:1 serial dilutions of your 0.2 mg/mL antibody.  Add 5 µL of each dilution (including the undiluted antibody) to 100 µL of cells (at 107 cells/ml) to test six concentrations - 1.0, 0.5, 0.25, 0.125, 0.06, and 0.03 µg per million cells in 100 µL volume. Compare staining patterns or create a titration curve using the MFI or staining index to determine the optimal concentration. 

I can’t find the antibody-dye combination that I need. When will it be available?

We continuously update our catalog, introducing scores of new products every month. Please get in touch with our Technical Service team for an update on new products or recommendations for suitable alternatives to complete your panel. Or contact Custom Solutions to inquire about our affordable custom conjugation services. 

I need help to validate the performance of my Flexi-Fluor. Who should I contact?

Please get in touch with Technical Service for assistance. 

Can I order more than 50 μg of a Flexi-Fluor?

Yes, you can order multiple vials of the same Flexi-Fluor products.  We cannot guarantee, however, that these vials will be bottled from the same lot. For bulk single-lot orders, contact our Custom Solutions team.

What is the expiration date of my Flexi-Fluor?

Expiration dates can be found on the vial label or by using our CoA lookup tool. 

Other Formats

View All CD20 Reagents Request Custom Conjugation
Description Clone Applications
APC anti-human CD20 2H7 FC
FITC anti-human CD20 2H7 FC
PE anti-human CD20 2H7 FC
PE/Cyanine5 anti-human CD20 2H7 FC
Purified anti-human CD20 2H7 FC,CyTOF®,IHC-F,IP
APC/Cyanine7 anti-human CD20 2H7 FC
PE/Cyanine7 anti-human CD20 2H7 FC
Alexa Fluor® 488 anti-human CD20 2H7 FC,IHC-F
Alexa Fluor® 647 anti-human CD20 2H7 FC,IHC-F
Pacific Blue™ anti-human CD20 2H7 FC
Alexa Fluor® 700 anti-human CD20 2H7 FC
PerCP anti-human CD20 2H7 FC
PerCP/Cyanine5.5 anti-human CD20 2H7 FC
Brilliant Violet 421™ anti-human CD20 2H7 FC,IHC-F
Brilliant Violet 570™ anti-human CD20 2H7 FC
Brilliant Violet 605™ anti-human CD20 2H7 FC
Brilliant Violet 650™ anti-human CD20 2H7 FC
Brilliant Violet 785™ anti-human CD20 2H7 FC
Brilliant Violet 510™ anti-human CD20 2H7 FC
Brilliant Violet 711™ anti-human CD20 2H7 FC
Purified anti-human CD20 (Maxpar® Ready) 2H7 FC,CyTOF®
PE/Dazzle™ 594 anti-human CD20 2H7 FC
Biotin anti-human CD20 2H7 FC
APC/Fire™ 750 anti-human CD20 2H7 FC
Alexa Fluor® 594 anti-human CD20 2H7 ICC,IHC-F
FITC anti-human CD20 2H7 FC
Pacific Blue™ anti-human CD20 2H7 FC
APC anti-human CD20 2H7 FC
PE/Cyanine7 anti-human CD20 2H7 FC
TotalSeq™-A0100 anti-human CD20 2H7 PG
TotalSeq™-B0100 anti-human CD20 2H7 PG
TotalSeq™-C0100 anti-human CD20 2H7 PG
PerCP/Cyanine5.5 anti-human CD20 2H7 FC
Spark NIR™ 685 anti-human CD20 2H7 FC
Spark YG™ 593 anti-human CD20 2H7 FC
GMP FITC anti-human CD20 2H7 FC
TotalSeq™-D0100 anti-human CD20 2H7 PG
GMP APC anti-human CD20 2H7 FC
Spark Violet™ 500 anti-human CD20 2H7 FC
GMP Pacific Blue™ anti-human CD20 2H7 FC
GMP PerCP/Cyanine5.5 anti-human CD20 2H7 FC
Spark Violet™ 538 anti-human CD20 2H7 FC
APC/Fire™ 750 anti-human CD20 2H7 FC
PE anti-human CD20 2H7 FC
GMP PE/Cyanine7 anti-human CD20 2H7 FC
Spark Blue™ 515 anti-human CD20 2H7 FC
GMP APC/Fire™ 750 anti-human CD20 2H7 FC
GMP PE anti-human CD20 2H7 FC
Spark Blue™ 550 anti-human CD20 (Flexi-Fluor™) 2H7 FC
Spark Blue™ 574 anti-human CD20 (Flexi-Fluor™) 2H7 FC
Spark Red™ 718 anti-human CD20 (Flexi-Fluor™) 2H7 FC
APC/Fire™ 810 anti-human CD20 2H7 FC
Alexa Fluor® 660 anti-human CD20 2H7 FC
Go To Top Version: 1    Revision Date: 02.06.2024

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

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

BioLegend, the BioLegend logo, and all other trademarks are property of BioLegend, Inc. or their respective owners, and all rights are reserved.

 

8999 BioLegend Way, San Diego, CA 92121 www.biolegend.com
Toll-Free Phone: 1-877-Bio-Legend (246-5343) Phone: (858) 768-5800 Fax: (877) 455-9587

This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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