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Anti-CD20 Antibody Picoband™ (monoclonal, 4D11)

Figure 1. Western blot analysis of CD20 using anti-CD20 antibody (M03780-5).
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 30 ug of sample under reducing conditions.
Lane 1: human Jurkat whole cell lysates,
Lane 2: human Raji whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-CD20 antigen affinity purified monoclonal antibody (Catalog # M03780-5) 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-mouse IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for CD20 at approximately 33 kDa. The expected band size for CD20 is at 33 kDa.
Figure 2. IHC analysis of CD20 using anti-CD20 antibody (M03780-5).
CD20 was detected in a paraffin-embedded section of human gastric carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-CD20 Antibody (M03780-5) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.
Figure 3. IHC analysis of CD20 using anti-CD20 antibody (M03780-5).
CD20 was detected in a paraffin-embedded section of human lienal rupture tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 µg/ml mouse anti-CD20 Antibody (M03780-5) overnight at 4°C. Peroxidase Conjugated Goat Anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Mouse IgG Super Vision Assay Kit (Catalog # SV0001) with DAB as the chromogen.
Figure 4. IF analysis of CD3E and CD20 using anti-CD3E antibody (PB9093) and anti-CD20 antibody (M03780-5).
CD3E and CD20 was detected in a paraffin-embedded section of human tonsil tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 5 µg/mL rabbit anti-CD3E antibody (PB9093) and mouse anti-CD20 antibody (M03780-5) overnight at 4°C. DyLight®550 Conjugated Goat Anti-Rabbit IgG (BA1135), DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IHC, IF
Primary Accession	P11836
Host	Mouse
Isotype	Mouse IgG2b
Reactivity	Human
Clonality	Monoclonal
Format	Lyophilized
Description	Anti-CD20 Antibody Picoband™ (monoclonal, 4D11) . Tested in IF, IHC, WB applications. This antibody reacts with Human.
Reconstitution	Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Additional Information
Gene ID	931
Other 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
Calculated MW	33077 Da
Application Details	Western blot, 0.25-0.5 µg/ml, Human Immunohistochemistry(Paraffin-embedded Section), 2-5 µg/ml, Human Immunofluorescence, 5 µg/ml, Human
Source	Eukaryota
Contents	Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.
Clone Names	Clone: 4D11
Immunogen	E.coli-derived human CD20 recombinant protein (Position: M1-D261). Human CD20 shares 75% amino acid (aa) sequence identity with mouse CD20.
Purification	Immunogen affinity purified.
Storage	At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

For Research Use Only. Not For Use In Diagnostic Procedures.

Protein Information
Name	MS4A1
Synonyms	CD20
Function	B-lymphocyte-specific membrane protein that plays a role in the regulation of cellular calcium influx necessary for the development, differentiation, and activation of B-lymphocytes (PubMed:12920111, PubMed:3925015, PubMed:7684739). Functions as a store-operated calcium (SOC) channel component promoting calcium influx after activation by the B-cell receptor/BCR (PubMed:12920111, PubMed:18474602, PubMed:7684739).
Cellular Location	Cell membrane; Multi-pass membrane protein. Cell membrane; Lipid-anchor. Note=Constitutively associated with membrane rafts.
Tissue Location	Expressed on B-cells.

Research Areas

Application Protocols

Provided below are standard protocols that you may find useful for product applications.

Western Blot Protocol

Blocking Peptides Protocol

Dot Blot Protocol

Immunohistochemistry Protocol

Immunofluorescence Protocol

Immunoprecipitation Protocol

Flow Cytomety Protocol

Cell Culture Protocol

BACKGROUND

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.

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¥ 3,367.00

Cat# ABO16261

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100 µg

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Anti-CD20 Antibody Picoband™ (monoclonal, 4D11)

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