CD20 Recombinant Rabbit Monoclonal Antibody [SY12-01]
cat.: ET1605-33
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Monkey
Applications: WB, IHC-P, IF-Tissue, IF-Cell, FC, IP
Clonality: Monoclonal
Clone number: SY12-01
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 33 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human CD20 aa 248-297 / 297.
Positive control: Raji cell lysate, Ramos cell lysate, Daudi cell lysate, human tonsil tissue, human spleen tissue, B-cell lymphoma tissue, Raji.
Subcellular location: Cell membrane.
Recommended Dilutions:
  WB
  IHC-P
  IF-Tissue
  IF-Cell
  FC
  IP
1:1,000-1:5,000
1:200
1:50
1:100
1:1,000
1-2μg/sample
Uniprot #: SwissProt: P11836 Human
Alternative names: APY ATOPY B lymphocyte antigen CD20 B Lymphocyte Cell Surface Antigen B1 B-lymphocyte antigen CD20 B-lymphocyte cell-surface antigen B1 B-lymphocyte surface antigen B1 B1 Bp 35 Bp35 CD 20 CD20 CD20 antigen CD20 receptor CD20_HUMAN CVID5 Fc epsilon receptor I beta chain Fc Fragment of IgE high affinity I receptor for beta polypeptide FCER1B High affinity immunoglobulin epsilon receptor subunit beta IgE Fc receptor subunit beta IGEL IGER IGHER LEU 16 LEU16 leukocyte surface antigen Leu 16 Leukocyte surface antigen Leu-16 Ly44 Membrane spanning 4 domains A1 Membrane spanning 4 domains subfamily A member 2 Membrane-spanning 4-domains subfamily A member 1 MS4A1 MS4A2 S7
Images
ET1605-33_1.jpg Fig1: Western blot analysis of CD20 on different lysates with Rabbit anti-CD20 antibody (ET1605-33) 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.

Predicted band size: 33 kDa
Observed band size: 33 kDa

Exposure time: 4 seconds; ECL: K1801;

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 (ET1605-33) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1605-33_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human tonsil tissue with Rabbit anti-CD20 antibody (ET1605-33) 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 (ET1605-33) 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.
ET1605-33_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human spleen 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 (ET1605-33, 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.
ET1605-33_4.jpg Fig4: 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 (ET1605-33, 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.
ET1605-33_5.jpg Fig5: Immunofluorescence analysis of paraffin-embedded human tonsil tissue labeling CD20 with Rabbit anti-CD20 antibody (ET1605-33) 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 (ET1605-33, green) at 1/50 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
ET1605-33_6.jpg Fig6: Immunocytochemistry analysis of Raji (positive) and 293T (negative) labeling CD20 with Rabbit anti-CD20 antibody (ET1605-33) 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 (ET1605-33) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1605-33_7.jpg Fig7: CD20 was immunoprecipitated from 0.2 mg Raji cell lysate with ET1605-33 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using ET1605-33 at 1/2,000 dilution. Mouse anti Rabbit IgG heavy chain (Fc) secondary antibody (M1003-7) at 1/10,000 dilution was used for 1 hour at room temperature.

Lane 1: Raji cell lysate (input)
Lane 2: ET1605-33 IP in Raji cell lysate
Lane 3: Rabbit IgG instead of ET1605-33 in Raji cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 10 seconds; ECL: K1801
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.