CD4 Overexpression Lysate

Name: CD4 Overexpression Lysate
Brand: Novus Biologicals
SKU: NBP2-04168

Novus Biologicals, part of Bio-Techne | Catalog # NBP2-04168

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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 CD4 Overexpression Lysate

CD4 Transient Overexpression Lysate
Expression Host: HEK293T

Plasmid: RC206453

Accession#: NM_000616

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

48.3 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 CD4 Overexpression Lysate

Western Blot: CD4 Overexpression Lysate [NBP2-04168]

Western Blot: CD4 Overexpression Lysate (Adult Normal) [NBP2-04168] Left-Empty vector transfected control cell lysate (HEK293 cell lysate); Right -Over-expression Lysate for CD4.

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: CD4

CD4 (cluster of differentiation 4), also known as L3T4 or T4, is a 55 kDa single chain type I transmembrane glycoprotein belonging to the immunoglobin (Ig) superfamily. CD4 is predominantly expressed on most thymocytes, a subset of mature T lymphocytes, and weakly on monocytes, tissue macrophages, dendritic cells, and granulocytes. It is also expressed on neurons and glial cells in the brain (1). CD4 is expressed along with CD8 on double positive T cells during their development in the thymus. Either CD4 or CD8 expression is then lost giving rise to single positive (SP) CD4+ or CD8+ mature T cells. CD4+ SP cells (T helper cells) further differentiate into multiple subsets of CD4+ cells including Th1, Th2, Th17, Tfh, and Treg cells which regulate humoral and cellular immunity (2). The extracellular region of CD4 consists of 372 amino acids (aa) with four immunoglobin-like domains (D1-D4). The structures of D1 and D3 resemble variable (IgV) domains while D2 and D4 resemble constant (IgC) domains (3).

Given its critical role in T cell development, CD4 also has diverse immunology-related functions. CD4 acts as a coreceptor with the T-cell receptor (TCR) during T cell activation and thymic differentiation by binding directly to major histocompatibility complex (MHC) class II antigens and associating with the protein tyrosine kinase, Lck (4). This interaction contributes to the formation of the immunological synapse (5). Defects in antigen presentation cause dysfunction of CD4+ T cells and the almost complete loss of MHC II expression on B cells in peripheral blood, as observed in severe combined immunodeficiency (SCID) (6). CD4 also functions as a receptor for the human immunodeficiency virus (HIV) by binding to gp120, the envelope glycoprotein of HIV-1. It has been shown that the V-like domains are critical for binding to gp120 (7). In immune mediated and infectious diseases of the central nervous system, CD4 functions as an indirect mediator of neuronal damage (8).

References

1. Omri, B., Crisanti, P., Alliot, F., Marty, M., Rutin, J., Levallois, C., . . . Pessac, B. (1994). CD4 expression in neurons of the central nervous system. International Immunology, 6(3), 377-385. doi:10.1093/intimm/6.3.377

2. Wan, Y. Y., & Flavell, R. A. (2009). How diverse-CD4 effector T cells and their functions. Journal of Molecular Cell Biology, 1(1), 20-36. doi:10.1093/jmcb/mjp001

3. Wu, H., Myszka, D. G., Tendian, S. W., Brouillette, C. G., Sweet, R. W., Chaiken, I. M., & Hendrickson, W. A. (1996). Kinetic and structural analysis of mutant CD4 receptors that are defective in HIV gp120 binding. Proceedings of the National Academy of Sciences, 93(26), 15030-15035. doi:10.1073/pnas.93.26.15030

4. Doyle, C., & Strominger, J. L. (1987). Interaction between CD4 and class II MHC molecules mediates cell adhesion. Nature, 330, 256-259. doi:10.1038/330256a0

5. Vignali, D. A. (2010). CD4 on the road to coreceptor status. The Journal of Immunology, 184(11), 5933-5934. doi:10.4049/jimmunol.1090037

6. Tasher, D., & Dalal, I. (2012). The genetic basis of severe combined immunodeficiency and its variants. The Application of Clinical Genetics, 5, 67-80. doi:10.2147/tacg.s18693

7. Arthos, J., Deen, K. C., Chaikin, M. A., Fornwald, J. A., Sathe, G., Sattentau, Q. J., . . . Sweet, R. W. (1989). Identification of the residues in human CD4 critical for the binding of HIV. Cell, 57(3), 469-481. doi:10.1016/0092-8674(89)90922-7

8. Buttini, M., Westland, C. E., Masliah, E., Yafeh, A. M., Wyss-Coray, T., Mucke, L. (1998). Novel role of human cd4 molecule identified in neurodegeneration. Nature Medicine, 4(4), 441-446. doi:10.1038/nm0498-441

Alternate Names

CD4

Gene Symbol

CD4

Additional CD4 Products

Product Documents for CD4 Overexpression Lysate

Product Datasheets

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COA

SDS

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Product Specific Notices for CD4 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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