Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)

Rabbit Recombinant Monoclonal CD20 antibody. Carrier free. Suitable for IP, WB, IHC-P, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 2 publications.

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Images

Western blot - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (AB166865), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	IHC-P	ICC/IF	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Rat	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended
Monkey	Not recommended	Not recommended	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes -
Species Monkey	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Monkey	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes For optimal performance in IHC, the primary antibody should be incubated overnight at 4℃. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes For optimal performance in IHC, the primary antibody should be incubated overnight at 4℃. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info -	Notes For optimal performance in IHC, the primary antibody should be incubated overnight at 4℃. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Monkey	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Monkey	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Monkey	Dilution info -	Notes -

Associated Products

Select an associated product type

15 products for Alternative Product

10 products for Alternative Version

Target data

See full target information MS4A1

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).

Alternative names

CD20, MS4A1, B-lymphocyte antigen CD20, B-lymphocyte surface antigen B1, Bp35, Leukocyte surface antigen Leu-16, Membrane-spanning 4-domains subfamily A member 1

Recommended products

Rabbit Recombinant Monoclonal CD20 antibody. Carrier free. Suitable for IP, WB, IHC-P, ICC/IF, Flow Cyt (Intra) and reacts with Human samples. Cited in 2 publications.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EP459Y
Purification technique: Affinity purification Protein A
Specificity: For optimal performance in IHC, the primary antibody should be incubated overnight at 4℃.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab166865 is the carrier-free version of Anti-CD20 antibody [EP459Y] ab78237.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

What does low endotoxin mean?
Our low endotoxin, azide-free formats have low endotoxin level (1 EU/mg, determined by the TAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

CD20 also known as MS4A1 or L26 is a membrane-spanning 4-domains subfamily A member 1 protein. It is a non-glycosylated phosphoprotein with a molecular mass of approximately 33-37 kDa. Expressed extensively on the surface of B-cells CD20 plays an important role in B-cell activation and regulation. While CD20 is absent on early pro-B cells and plasma cells its presence increases as B-cells mature. Anti-CD20 antibodies such as 2H7 are commonly used in research and treatment to deplete B-cells due to the target's consistent expression in most stages of B-cell development.

Biological function summary

CD20 contributes significantly to calcium ion transport which is essential for the activation process of B-cells. Although CD20 does not belong to a larger protein complex its role centers around forming a calcium channel that allows a sustained influx of calcium into the B-cells. This influx is important for the signaling pathways that govern B-cell activation proliferation and differentiation impacting the immune response efficacy.

Pathways

CD20 is closely involved in the B-cell receptor (BCR) signaling pathway and the Fc gamma R-mediated phagocytosis pathway. These pathways play vital roles in adaptive immunity and immune response modulation. CD20's interaction with proteins like PI3K during BCR signaling enhances receptor-mediated cellular signals subsequently influencing downstream effectors involved in cell growth and survival of B-cells.

Associated diseases and disorders

CD20 is strongly linked to certain blood cancers and autoimmune diseases including non-Hodgkin's lymphoma and rheumatoid arthritis. These conditions often exploit aberrant B-cell activity or count. CD20's expression on malignant B-cells in non-Hodgkin's lymphoma makes it an effective target for monoclonal antibody therapies such as rituximab an anti-CD20 antibody. Additionally CD20-targeted therapies can help deplete pathogenic B-cells in rheumatoid arthritis highlighting the protein's relevance in disease treatment.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

15 product images

Western blot - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)
False colour image of Western blot: Anti-CD20 antibody [EP459Y] – Mouse IgG2a (Chimeric) staining at 1/1000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (Anti-alpha Tubulin antibody [EP1332Y] - Loading Control ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-CD20 antibody [EP459Y] - Mouse IgG2a (Chimeric) ab279299 was shown to bind specifically to CD20. A band was observed at 33 kDa in wild-type Raji cell lysates with no signal observed at this size in MS4A1 knockout cell line Human MS4A1 (CD20) knockout Raji cell line ab273871 (knockout cell lysate Human MRPS28 knockout HEK-293T cell lysate ab263259). To generate this image, wild-type and MS4A1 knockout Raji cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1% Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.
Lanes 1 - 4: Western blot - Anti-CD20 antibody [EP459Y] (Anti-CD20 antibody [EP459Y] ab78237) at 1/1000 dilution
Lanes 1 - 4: Western blot - Anti-CD20 antibody [EP459Y] - Mouse IgG2a (Chimeric) (Anti-CD20 antibody [EP459Y] - Mouse IgG2a (Chimeric) ab279299) at 1/1000 dilution
Lane 1: Wild-type Raji cell lysate at 20 µg
Lane 2: MS4A1 knockout Raji cell lysate at 20 µg
Lane 2: Western blot - Human MS4A1 (CD20) knockout Raji cell line (Human MS4A1 (CD20) knockout Raji cell line ab273871)
Lane 3: A549 cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 21 kDa, 33 kDa
Observed band size: 33 kDa
Western blot - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)
False colour image of Western blot: Anti-CD20 antibody [EP459Y] - Rat IgG2a (Chimeric) staining at 1/1000 dilution, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (Anti-alpha Tubulin antibody [EP1332Y] - Loading Control ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-CD20 antibody [EP459Y] - Rat IgG2a (Chimeric) ab279300 was shown to bind specifically to CD20. A band was observed at 33 kDa in wild-type Raji cell lysates with no signal observed at this size in MS4A1 knockout cell line Human MS4A1 (CD20) knockout Raji cell line ab273871 (knockout cell lysate Human MRPS28 knockout HEK-293T cell lysate ab263259). To generate this image, wild-type and MS4A1 knockout Raji cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1% Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rat IgG H&L (IRDye^® 800CW) preabsorbed (ab253031) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.
Lanes 1 - 4: Western blot - Anti-CD20 antibody [EP459Y] (Anti-CD20 antibody [EP459Y] ab78237) at 1/1000 dilution
Lanes 1 - 4: Western blot - Anti-CD20 antibody [EP459Y] - Rat IgG2a (Chimeric) (Anti-CD20 antibody [EP459Y] - Rat IgG2a (Chimeric) ab279300) at 1/1000 dilution
Lane 1: Wild-type Raji cell lysate at 20 µg
Lane 2: MS4A1 knockout Raji cell lysate at 20 µg
Lane 2: Western blot - Human MS4A1 (CD20) knockout Raji cell line (Human MS4A1 (CD20) knockout Raji cell line ab273871)
Lane 3: A549 cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 21 kDa, 33 kDa
Observed band size: 33 kDa
Immunoprecipitation - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)
Anti-CD20 antibody [EP459Y] ab78237 (purified) at 1/20 immunoprecipitating CD20 in 10 μg Ramos cell lysate (Lanes 1 and 2, observed at 33 kDa). Lane 3 - Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730). For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10,000 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD20 antibody [EP459Y] ab78237).
All lanes: Immunoprecipitation - Anti-CD20 antibody [EP459Y] (Anti-CD20 antibody [EP459Y] ab78237)
Predicted band size: 33 kDa
Immunocytochemistry/ Immunofluorescence - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)
Immunocytochemistry/ Immunofluorescence analysis of Ramos (human Burkitt's lymphoma B lymphocyte) labeling CD20 with purified Anti-CD20 antibody [EP459Y] ab78237 at 1/10 dilution. Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 was used as the secondary antibody. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200. PBS instead of the primary antibody was used as negative control. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Nuclei were stained with DAPI (blue).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD20 antibody [EP459Y] ab78237).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)
This IHC data was generated using the same anti-CD20 antibody clone, EP459Y, in a different buffer formulation (cat# Anti-CD20 antibody [EP459Y] ab78237).
Immunohistochemical staining of paraffin embedded human B cell lymphoma with purified Anti-CD20 antibody [EP459Y] ab78237 at a working dilution of 1/50. The secondary antibody used is Goat Anti-Rabbit IgG H&L (HRP) ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
Flow Cytometry (Intracellular) - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)
Intracellular Flow Cytometry analysis ofRamos (Human Burkitt's lymphoma B lymphocyte) labeling CD20 with purified Anti-CD20 antibody [EP459Y] ab78237 at 1/200 dilution (Red). Goat anti rabbit IgG (Alexa Fluor^®488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as secondary antibody. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol.
Isotype control: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black)
Unlabelled cells: (Blue)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD20 antibody [EP459Y] ab78237).
Western blot - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)
False colour image of Western blot: Anti-CD20 antibody [EP459Y] - Mouse IgG1 (Chimeric) staining, shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (Anti-alpha Tubulin antibody [EP1332Y] - Loading Control ab52866) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-CD20 antibody [EP459Y] - Mouse IgG1 (Chimeric) ab279298 was shown to bind specifically to CD20. A band was observed at 33 kDa in wild-type Raji cell lysates with no signal observed at this size in MS4A1 knockout cell line Human MS4A1 (CD20) knockout Raji cell line ab273871 (knockout cell lysate Human MRPS28 knockout HEK-293T cell lysate ab263259). To generate this image, wild-type and MS4A1 knockout Raji cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) at 1/20000 dilution.
Lanes 1 - 4: Western blot - Anti-CD20 antibody [EP459Y] - Mouse IgG1 (Chimeric) (Anti-CD20 antibody [EP459Y] - Mouse IgG1 (Chimeric) ab279298) at 1/1000 dilution
Lanes 1 - 4: Western blot - Anti-CD20 antibody [EP459Y] (Anti-CD20 antibody [EP459Y] ab78237) at 1/1000 dilution
Lane 1: Wild-type Raji cell lysate at 20 µg
Lane 2: MS4A1 knockout Raji cell lysate at 20 µg
Lane 2: Western blot - Human MS4A1 (CD20) knockout Raji cell line (Human MS4A1 (CD20) knockout Raji cell line ab273871)
Lane 3: A549 cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 21 kDa, 33 kDa
Observed band size: 33 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)
Unpurified Anti-CD20 antibody [EP459Y] ab78237 staining human CD20 in human lymphoma tissue by immunohistochemistry using formalin fixed, paraffin embedded tissue.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD20 antibody [EP459Y] ab78237).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)
Unpurified Anti-CD20 antibody [EP459Y] ab78237 showing positive staining in normal human spleen tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD20 antibody [EP459Y] ab78237).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)
Unpurified Anti-CD20 antibody [EP459Y] ab78237 showing negative staining in normal human brain tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD20 antibody [EP459Y] ab78237).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)
Unpurified Anti-CD20 antibody [EP459Y] ab78237 showing negative staining in normal human heart tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD20 antibody [EP459Y] ab78237).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)
Unpurified Anti-CD20 antibody [EP459Y] ab78237 showing negative staining in normal human kidney tissue.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD20 antibody [EP459Y] ab78237).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)
This IHC data was generated using the same anti-CD20 antibody clone, EP459Y, in a different buffer formulation (cat# Anti-CD20 antibody [EP459Y] ab78237).
Unpurified Anti-CD20 antibody [EP459Y] ab78237 showing positive staining in normal human tonsil tissue.
Flow Cytometry (Intracellular) - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD20 antibody [EP459Y] ab78237).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD20 antibody [EP459Y] - Low endotoxin, Azide free (ab166865)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-CD20 antibody [EP459Y] ab78237).
Immunohistochemical analysis of formalin fixed paraffin embedded human spleen labelling CD20 with Anti-CD20 antibody [EP459Y] ab78237 at a concentration of 1 µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH 8.5 for 32mins.
Anti-CD20 antibody [EP459Y] ab78237 anti-CD20 [EP459Y] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).