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

Product Name

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

View all CD20 Antibodies

SKU/Catalog Number

M03780-5

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-CD20 Antibody Picoband™ (monoclonal, 4D11) catalog # M03780-5. Tested in IF, IHC, WB applications. This antibody reacts with Human.

Storage & Handling

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.

Cite This Product

Anti-CD20 Antibody Picoband™ (monoclonal, 4D11) (Boster Biological Technology, Pleasanton CA, USA, Catalog # M03780-5)

Host

Mouse

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl and 0.2 mg Na2HPO4.

Clonality

Monoclonal

Clone Number

4D11

Isotype

Mouse IgG2b

Immunogen

E.coli-derived human CD20 recombinant protein (Position: M1-D261). Human CD20 shares 75% amino acid (aa) sequence identity with mouse CD20.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross-reactivity with other proteins.

Reactive Species

M03780-5 is reactive to MS4A1 in Human

Applications

M03780-5 is guaranteed for IF, IHC, WB Boster Guarantee

Observed Molecular Weight

33 kDa

Calculated molecular weight

33.077kDa

Background of 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.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 μg/ml, Human
Immunohistochemistry(Paraffin-embedded Section), 2-5 μg/ml, Human
Immunofluorescence, 5 μg/ml, Human

Validation Images & Assay Conditions

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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.

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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.

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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.

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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.

Gene/Protein Information For MS4A1 (Source: Uniprot.org, NCBI)

Alternative Names

B1; B-lymphocyte antigen CD20; B-lymphocyte cell-surface antigen B1; B-lymphocyte surface antigen B1; Bp35; Bp35MGC3969; CD20 antigen; CD20 receptor; CD20; CD20S7; CVID5; LEU-16; Leukocyte surface antigen Leu-16; Ly-44; Membrane-spanning 4-domains subfamily A member 1; membrane-spanning 4-domains, subfamily A, member 1; MS4A1; MS4A2; S7 MS4A1 B1, Bp35, CD20, CVID5, FMC7, LEU-16, MS4A2, S7 membrane spanning 4-domains A1 B-lymphocyte antigen CD20|B-lymphocyte cell-surface antigen B1|CD20 antigen|CD20 receptor|leukocyte surface antigen Leu-16|membrane-spanning 4-domains, subfamily A, member 1

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