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SKU:PB9050
Clonality:Polyclonal
Application:Flow Cytometry, IHC-P, IHC-F, ICC, WB
Price: $240.00
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Overview

Product Name Anti-CD20 Picoband™ Antibody
SKU/Catalog Number PB9050
Description Rabbit IgG polyclonal antibody for B-lymphocyte antigen CD20(MS4A1) detection. Tested with WB, IHC-P, IHC-F, ICC, FCM in Human.
Cite This Product Anti-CD20 Picoband™ Antibody (Boster Biological Technology, Pleasanton CA, USA, Catalog # PB9050)
Replacement Item This antibody may replace the following items: sc-134340|sc-15361|sc-19990|sc-292152|sc-33488|sc-33489|sc-33490|sc-33491|sc-33493|sc-393789|sc-393894|sc-51531|sc-58982|sc-58983|sc-58985|sc-70582|sc-7733|sc-7735|sc-82536 from Santa Cruz Biotechnology.
Host Rabbit
Isotype N/A
Validated Species Human
Application Flow Cytometry, IHC-P, IHC-F, ICC, WB

*Our Boster Guarantee covers the use of this product in the above tested applications.

**For positive and negative control design, consult "Tissue specificity" under Protein Target Info.

Recommended Detection Systems Boster recommends Enhanced Chemiluminescent Kit with anti-Rabbit IgG (EK1002) for Western blot, and HRP Conjugated anti-Rabbit IgG Super Vision Assay Kit (SV0002-1) for IHC(P), IHC(F) and ICC.
*Blocking peptide can be purchased at $50. Contact us for more information
**Boster also offers various secondary antibodies for Immunoflourescecne and IHC. Take advantage of the buy 1 primary antibody get 1 secondary antibody for free promotion for the entire year 2017!
Immunogen E.coli-derived human CD20 recombinant protein (Position: M1-D261). Human CD20 shares 75% amino acid (aa) sequence identity with mouse CD20.
Cross Reactivity No cross reactivity with other proteins
Pack Size 100μg/vial

Properties

Clonality Polyclonal
Form Lyophilized
Contents Each vial contains 5mg BSA, 0.9mg NaCl, 0.2mg Na2HPO4, 0.05mg NaN3.
*carrier free antibody available upon request.
Concentration Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
Storage At -20˚C for one year. After reconstitution, at 4˚C for one month. It can also be aliquotted and stored frozen at -20˚C for a longer time.Avoid repeated freezing and thawing.
Purification Immunogen affinity purified.
Isotype N/A

Protein Target Info (Source: Uniprot.org)

You can check the tissue specificity below for information on selecting positive and negative control.

Gene Name MS4A1
Protein Name B-lymphocyte antigen CD20
Molecular Weight 33077 MW
Protein Function This protein may be involved in the regulation of B-cell activation and proliferation.
Tissue Specificity Expressed on B-cells.
Sequence Similarities Belongs to the MS4A family.
Subcellular Localization Cell membrane ; Multi-pass membrane protein . Cell membrane ; Lipid-anchor .
Uniprot ID P11836
Alternative Names B-lymphocyte antigen CD20;B-lymphocyte surface antigen B1;Bp35;Leukocyte surface antigen Leu-16;Membrane-spanning 4-domains subfamily A member 1;CD20;MS4A1;CD20;
Research Areas |signal transduction|adapters|transmembrane|
*if product is indicated to react with multiple species, protein info is based on the human gene.

Background for B-lymphocyte antigen CD20

CD20, also known as MS4A1, is an activated-glycosylated phosphoprotein expressed on the surface of all B-cells beginning at the pro-B phase (CD45R+, CD117+) and progressively increasing in concentration until maturity. It is mapped to 11q12.2. This gene encodes a member of the membrane-spanning 4A gene family. The function of CD20 is to enable optimal B-cell immune response, specifically against T-independent antigens. It is suspected that CD20 acts as a calcium channel in the cell membrane. What’s more, this protein may be involved in the regulation of B-cell activation and proliferation.

Anti-CD20 Picoband™ Antibody Images

Click the images to enlarge.

Anti-CD20 Picoband™ Antibody
Figure 1. IHC analysis of CD20 using anti-CD20 antibody (PB9050).
CD20 was detected in paraffin-embedded section of Human Tonsil Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CD20 Antibody (PB9050) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Anti-CD20 Picoband™ Antibody
Figure 2. IHC analysis of CD20 using anti-CD20 antibody (PB9050).
CD20 was detected in paraffin-embedded section of Human Tonsil Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-CD20 Antibody (PB9050) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.
Anti-CD20 Picoband™ Antibody
Figure 3. Western blot analysis of CD20 using anti-CD20 antibody (PB9050).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: RAJI Whole Cell Lysate
Lane 2: CEM Whole Cell Lysate
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-CD20 antigen affinity purified polyclonal antibody (Catalog # PB9050) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for CD20 at approximately 33KD. The expected band size for CD20 is at 33KD.
Anti-CD20 Picoband™ Antibody
Figure 4. Flow Cytometry analysis of U937 cells using anti-CD20 antibody (PB9050).
Overlay histogram showing U937 cells stained with PB9050 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CD20 Antibody (PB9050,1μg/1x106 cells) for 30 min at 20°C. DyLight?488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
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Publications

Ni C, Chen Y, Zeng M, Pei R, Du Y, Tang L, Wang M, Hu Y, Zhu H, He M, Wei X, Wang S, Ning X, Wang M, Wang J, Ma L, Chen X, Sun Q, Tang H, Wang Y, Wang X. Cell Res. 2015 Jul;25(7):785-800. Doi: 10.1038/Cr.2015.50. Epub 2015 Apr 28. In-Cell Infectio...

FAQs

Q: Do you offer BSA-free antibodies? Keyword: Bovine serum albumin, carrier protein, conjugation
A: Yes, please contact us at support@bosterbio.com for more information about BSA-free antibodies and availability. The new BSA-free formula uses trehalose as a replacement to BSA. We have tested many alternative chemicals and found that trehalose protects the antibodies the best.
Q: Can I conjugate markers to this antibody? Can I link custom conjugates to this antibody? Keyword: conjugation
A: The antibody is stored with BSA and cannot be conjugated with markers. Carrier free antibodies are available upon request. Please contact support@bosterbio.com
Q: What should I use for negative control?
A: Please contact us for negative control suggestions. You can also check expression databases such as genecards, uniprot etc. Due to logistic reasons, we do not sell serum or lysates that we use internally for positive or negative control.
Q: Where can I find troubleshooting information? What should I do if I have unexpected bands, high background, no signal, weak signal
A: You can find Boster's troubleshoot guides under tech support tab. Please contact us for further assistance on troubleshooting your experiment.
Q: What is the immunogen sequence of this antibody? Is this antibody polyclonal or monoclonal?
A: You can find the immunogen sequence under "Immunogen" and clonality in the product name.
Q: What is the expected band size? Why is it different than the observed band size?
A: The expected band size is predicted on the size of the protein. The actual band size may be affected by a few other factors including but not limited to:
1. Post-translational modification:phosphorylation, methylation, glycosylation etc. These modifications prevent SDS molecules from binding to the target protein and thus make the band size appear larger than expected
2. Post-translational cleavage: this can cause smaller bands and or multiple bands

3. Alternative splicing: the same gene can have alternative splicing patterns generating different size proteins, all with reactivities to the antibody.

4. Amino Acid R chain charge: SDS binds to positive charges. The different size and charge of the Amino Acid side chains can affect the amount of SDS binding and thus affect the observed band size.
5. Multimers: Multimers are usually broken up in reducing conditions. However if the interactions between the multimers are strong, the band may appear higher.,
Q: What is the suggested dilution ratio for Western Blot (WB), Immunohistochemistry (IHC) and or ELISA standards? What is the optimal pH for the sample?
A: Check the datasheet for the product for details on dilution ratios for different experiments. You can find the datasheet button on the right side of the product page.
Q: What is the protocol you used for your Western blotting (WB) and Immunohistochemistry (IHC)?
A: Check our protocols under the tech support tab.
Q: What are some alternative names that could be used to describe this product?
A: Some common names include but are not limited to cd20 antibody, b1 antibody, cd 20 antibody, fceri antibody, ms4a1 antibody