WB
Western blot analysis of CD20 on different lysates with Rabbit anti-CD20 antibody at 1/2,000 dilution. Lane 1: Raji cell lysate, Lane 2: 293T cell lysate (negative), Lane 3: Ramos cell lysate, Lane 4: HeLa cell lysate (negative), Lane 5: Daudi cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 4 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-CD20 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded B-cell lymphoma tissue using anti-CD20 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.ICC/IF
Immunocytochemistry analysis of Raji (positive) and 293T (negative) labeling CD20 with Rabbit anti-CD20 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD20 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.IF-P
Immunofluorescence analysis of paraffin-embedded human tonsil tissue labeling CD20 with Rabbit anti-CD20 antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (green) at 1/50 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).IP
CD20 was immunoprecipitated from 0.2 mg Raji cell lysate with Rabbit anti-CD20 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-CD20 antibody at 1/2,000 dilution. Mouse anti Rabbit IgG heavy chain (Fc) secondary antibody at 1/10,000 dilution was used for 1 hour at room temperature. Lane 1: Raji cell lysate (input), Lane 2: Rabbit anti-CD20 antibody IP in Raji cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-CD20 antibody in Raji cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 10 seconds.| Product Name | CD20 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic peptide within Human CD20 aa 248-297 / 297. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | ICC/IFIF-PIHCIPWB |
| WB:1:1000-1:5000 IHC:1:200 ICC/IF:1:100 IF-P:1:50 IP:1-2μg/sample |
|
| Species Reactivity | Human |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | MS4A1 |
|---|---|
| Gene Synonyms | B1 S7 Bp35 CD20 FMC7 CVID5 LEU-16 |
| Gene Full Name | membrane spanning 4-domains A1 |
| Gene Summary | This gene encodes a member of the membrane-spanning 4A gene family. Members of this nascent protein family are characterized by common structural features and similar intron/exon splice boundaries and display unique expression patterns among hematopoietic cells and nonlymphoid tissues. This gene encodes a B-lymphocyte surface molecule which plays a role in the development and differentiation of B-cells into plasma cells. This family member is localized to 11q12, among a cluster of family members. Alternative splicing of this gene results in two transcript variants which encode the same protein. [provided by RefSeq, Jul 2008] |
| Molecular Weight(MW) | 33kDa |
| Cellular Localization | Cell membrane. |
WB
Western blot analysis of CD20 on different lysates with Rabbit anti-CD20 antibody at 1/2,000 dilution. Lane 1: Raji cell lysate, Lane 2: 293T cell lysate (negative), Lane 3: Ramos cell lysate, Lane 4: HeLa cell lysate (negative), Lane 5: Daudi cell lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 4 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-CD20 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded B-cell lymphoma tissue using anti-CD20 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ICC/IF
Immunocytochemistry analysis of Raji (positive) and 293T (negative) labeling CD20 with Rabbit anti-CD20 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-CD20 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
IF-P
Immunofluorescence analysis of paraffin-embedded human tonsil tissue labeling CD20 with Rabbit anti-CD20 antibody at 1/50 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (green) at 1/50 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
IP
CD20 was immunoprecipitated from 0.2 mg Raji cell lysate with Rabbit anti-CD20 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-CD20 antibody at 1/2,000 dilution. Mouse anti Rabbit IgG heavy chain (Fc) secondary antibody at 1/10,000 dilution was used for 1 hour at room temperature. Lane 1: Raji cell lysate (input), Lane 2: Rabbit anti-CD20 antibody IP in Raji cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-CD20 antibody in Raji cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 10 seconds.| Application Notes | WB:1:1000-1:5000 IHC:1:200 ICC/IF:1:100 IF-P:1:50 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
Data sheet for OM644341