CD20 Overexpression Lysate

Novus Biologicals | Catalog # NBL1-13312

Novus Biologicals
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Key Product Details

Species

Human

Applications

Western Blot

Product Summary for CD20 Overexpression Lysate

CD20 Transient Overexpression Lysate


Expression Host: HEK293T

Plasmid: RC221570

Accession#: NM_152866

Protein Tag: C-MYC/DDK

You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT.

Product Specifications

Application Notes

This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct.

Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.

Theoretical MW

32.9 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Type

Overexpression

Scientific Data Images for CD20 Overexpression Lysate

Western Blot: CD20 Overexpression Lysate [NBL1-13312]

Western Blot: CD20 Overexpression Lysate [NBL1-13312]

Western Blot: CD20 Overexpression Lysate (Native) [NBL1-13312] - Detection of CD20 in lymphoma cell line lysates. CD20 over expression lysate was used as the positive control. This image was submitted via customer Review.
Western Blot: CD20 Overexpression Lysate [NBL1-13312]

Western Blot: CD20 Overexpression Lysate [NBL1-13312]

Western Blot: CD20 Overexpression Lysate (Native) [NBL1-13312] - Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CD20.

Formulation, Preparation, and Storage

Formulation

RIPA buffer

Concentration

The exact concentration of the protein of interest cannot be determined for overexpression lysates. Please contact technical support for more information.

Shipping

The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -80C. Avoid freeze-thaw cycles.

Background: CD20

CD20 is a non-glycosylated phosphoprotein that is expressed on the surface of normal and malignant B cells and functions in mediating calcium transport and B cell differentiation (1,2). CD20 is encoded by the membrane-spanning 4-domain family A member 1 (MS4A1) gene and, in humans, is located on chromosome 11q12 (1). The CD20 protein is 297 amino acids (aa) in length with a theoretical molecular weight (MW) of 33 kDa (1,2). Structurally, the CD20 protein has four membrane-spanning domains, two extracellular loop domains, and intracellular N- and C-terminal domains (1,2). CD20 is expressed at specific stages of B cell maturation including pre-B cells, mature naive and activated B cells, and memory B cells, but is absent from plasmablasts and plasma cells (1,3,4). Expression of CD20 is often increased on malignant B cells associated with various B cell disorders such as chronic lymphocytic leukemia (CLL), multiple sclerosis (MS), and rheumatoid arthritis (2-4). Anti-CD20 monoclonal antibody (mAb)-based therapies have become an appealing target for treating these immune-related disorders and cancers (1-5). Rituximab, a chimeric mAb, was the first FDA approved CD20 monoclonal antibody for the treatment of non-Hodgkin's lymphoma that is now commonly used to treat MS (2,3). Since its initial approval in 1997, several other chimeric and humanized anti-CD20 mAbs have been developed including Ofatumumab, Ublituximab, and Obinutuzumab (1-5). B cell depletion via CD20 mAbs can occur under different mechanisms such as antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), complement-dependent cytotoxicity (CDC), and direct induction of apoptosis (1-4). Many ongoing studies are focused on combination therapies with CD20 mAbs as an addition to chemotherapy, B cell receptor (BCR) signaling inhibitors, or BH3 mimetics (1,2,4). Additionally, the effects of bispecific antibodies and CD20 chimeric antigen receptor (CAR) T cell therapies are under investigation for the treatment of B cell malignancies (4,5).

References

1. Pavlasova G, Mraz M. The regulation and function of CD20: an "enigma" of B-cell biology and targeted therapy. Haematologica. 2020; 105(6):1494-1506. https://doi.org/10.3324/haematol.2019.243543

2. Payandeh Z, Bahrami AA, Hoseinpoor R, et al. The applications of anti-CD20 antibodies to treat various B cells disorders. Biomed Pharmacother. 2019; 109:2415-2426. https://doi.org/10.1016/j.biopha.2018.11.121

3. Margoni M, Preziosa P, Filippi M, Rocca MA. Anti-CD20 therapies for multiple sclerosis: current status and future perspectives. J Neurol. 2022; 269(3):1316-1334. https://doi.org/10.1007/s00415-021-10744-x

4. Klein C, Jamois C, Nielsen T. Anti-CD20 treatment for B-cell malignancies: current status and future directions. Expert Opin Biol Ther. 2021; 21(2):161-181. https://doi.org/10.1080/14712598.2020.1822318

5. Sharman JP. Targeting CD20: teaching an old dog new tricks. Hematology Am Soc Hematol Educ Program. 2019; 2019(1):273-278. https://doi.org/10.1182/hematology.2019000031

Long Name

Cluster of Differentiation 20

Alternate Names

B1, Bp35, CD20, LEU-16, Ly-44, MS4A1, S7

Gene Symbol

MS4A1

Additional CD20 Products

Product Documents for CD20 Overexpression Lysate

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for CD20 Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

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  • MS4A1/CD20 Overexpression Lysate (Native)
    Name: Christopher England
    Verified Customer | Posted 02/24/2017
    Detection of CD20 in lymphoma cell line lysates. CD20 over expression lysate was used as the positive control.
    Loaded 5 microliter into last well as positive control. In the future, we will load less protein. Overall, it looks great!
    CD20 Overexpression Lysate NBL1-13312

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