CD2 Overexpression Lysate

Name: CD2 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-08916

Novus Biologicals, part of Bio-Techne | Catalog # NBL1-08916

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Product Summary
Product Specifications
Scientific Data
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Species

Human

Applications

Western Blot

Product Summary for CD2 Overexpression Lysate

CD2 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC206612

Accession#: NM_001767

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.

TMW

39.3 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 CD2 Overexpression Lysate

Western Blot: CD2 Overexpression Lysate [NBL1-08916]

Western Blot: CD2 Overexpression Lysate (Adult Normal) [NBL1-08916] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CD2.

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: CD2

CD2, also known as sheep red blood cell receptor (SRBC-R), erythrocyte receptor, LFA-2, and T11, is a type I transmembrane glycoprotein that is expressed on the surface of T cells, natural killer (NK) cells, thymocytes, and dendritic cells (1,2). CD2 is a member of the immunoglobulin (Ig) superfamily and a costimulatory receptor that functions in formation of the immunological synapse and T cell activation and signaling (1). The human CD2 protein is 351 amino acids in length with a theoretical molecular weight of ~40 kDa, but a fully glycosylated protein can weight closer to 50 kDa (1,3). The CD2 protein contains a signal sequence, an extracellular domain (ECD) composed of an Ig-like V-type domain followed by an Ig-like C-type domain, a transmembrane helical domain, and a proline-rich cytoplasmic tail (1,3). CD2 binds with CD58, also called LFA-3, which is a surface glycoprotein expressed by antigen presenting cells (APCs) and other target cells (1,2). While CD58 is the primary ligand for CD2 in humans, it also interacts with CD59 and CD48, albeit with lower affinity (1,2). However, in mice and rats which lack CD58 the main ligand for CD2 is CD48 (4). Research has found that when there is no direct interaction, CD2 co-immunoprecipitates with the T cell receptor (TCR)/CD3 complex (1). CD2 is an adhesion molecule with a variety of functions including actin cytoskeleton rearrangement, immunological synapse formation through T cell-APC binding, thymocyte development and T cell activation, and NK cell activation (1,2). The immunological synapse forms upon T cell-APC engagement and creates a contact zone of supramolecule activation clusters (SMACs) where CD2-CD58 is part of the central SMAC (cSMAC) (2).

The CD2-CD58 interaction has been shown to play a role in anti-tumor immune response, where reduced CD58 signaling is associated with immune escape of tumor cells in various hematological and lymphoid malignancies, but restoration of the signal promotes an anti-tumor response (2,5). Additionally, following cytomegalovirus (CMV) infection, CD2's binding to upregulated CD58 on CMV-infected cells is crucial for the activation and function of adaptive NK cells in the anti-viral response (2). In contrast, in situations where there is an increase in T cell and NK cell activation, like various autoimmune disorders or following organ transplant, costimulatory blockade of CD2-CD58 may be a potential therapeutic treatment approach (1). Mouse and rat xenograft models have shown that blocking the CD2 using anti-CD2 monoclonal antibodies contributes to graft survival and protects against lymphocyte infiltration and inflammatory damage (2).

References

1. Binder C, Cvetkovski F, Sellberg F, et al. CD2 Immunobiology. Front Immunol. 2020;11:1090. https://doi.org/10.3389/fimmu.2020.01090

2. Zhang Y, Liu Q, Yang S, Liao Q. CD58 Immunobiology at a Glance. Front Immunol. 2021;12:705260. https://doi.org/10.3389/fimmu.2021.705260

3. Uniprot (P06729)

4. van der Merwe PA. A subtle role for CD2 in T cell antigen recognition. J Exp Med. 1999;190(10):1371-1374. https://doi.org/10.1084/jem.190.10.1371

5. Nishikori M, Kitawaki T, Tashima M, Shimazu Y, Mori M, et al. Diminished CD2 Expression in T cells Permits Tumor Immune Escape. J Clin Cell Immunol. 2016;7:406. https://doi.org/10.4172/2155-9899.1000406

Alternate Names

CD2, LFA-2, T11

Gene Symbol

CD2

Additional CD2 Products

Product Documents for CD2 Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for CD2 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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