Mouse CD86 HEK293 Overexpression Lysate

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Mouse CD86 HEK293 Overexpression Lysate: Product Information

Product Description
This Mouse CD86 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD86 protein (Cat: 50068-M02H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Mouse
Sequence Information
A DNA sequence encoding the mouse CD86 (NP_062261.3) (Met1-Glu245) was expressed with the Fc region of human IgG1 at the C-terminus.
Molecule Mass
The recombinant mouse CD86 consists of 460 amino acids and predicts a molecular mass of 52 kDa.

Mouse CD86 HEK293 Overexpression Lysate: Usage Guide

Preparation Method
Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube. 2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min. 3. Store the lyophilized cell lysate at 4℃. After re-dissolution, recommend to aliquot it into smaller quantities and store at -80℃.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Mouse CD86 HEK293 Overexpression Lysate: Alternative Names

Mouse B7 Overexpression Lysate; Mouse B7-2 Overexpression Lysate; Mouse B7.2 Overexpression Lysate; Mouse B70 Overexpression Lysate; Mouse Cd28l2 Overexpression Lysate; Mouse CLS1 Overexpression Lysate; Mouse ETC-1 Overexpression Lysate; Mouse Ly-58 Overexpression Lysate; Mouse Ly58 Overexpression Lysate; Mouse MB7 Overexpression Lysate; Mouse MB7-2 Overexpression Lysate; Mouse TS/A-2 Overexpression Lysate

CD86 Background Information

CD86, also known as B-lymphocyte activation antigen B7-2 (referred to as B7), is a member of the cell surface immunoglobulin superfamily. B7-2 exists predominantly as a monomer on cell surfaces and interacts with two co-stimulatory receptors CD28 and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) expressed on T cells, and thus induces the signal pathways which regulate T cell activation and tolerance, cytokine production, and the generation of CTL. It is indicated that contacts between B and T helper cells mediated by CD86 encourage signals for the proliferation and IgG secretion of normal B cells and B cell lymphomas. Recent study has revealed that CD86 also promotes the generation of a mature APC repertoire and promotes APC function and survival. CD86 has an important role in chronic hemodialysis, allergic pulmonary inflammation, arthritis, and antiviral responses, and thus is regarded as a promising candidate for immune therapy.
Full Name
CD86 antigen
References
  • Chen YQ, et al. (2006) CD28/CTLA-4--CD80/CD86 and ICOS--B7RP-1 costimulatory pathway in bronchial asthma. Allergy. 61(1): 15-26.
  • Rau FC, et al. (2009) B7-1/2 (CD80/CD86) direct signaling to B cells enhances IgG secretion. J Immunol. 183(12): 7661-71.
  • Dai ZS, et al. (2009) Defective expression and modulation of B7-2/CD86 on B cells in B cell chronic lymphocytic leukemia. Int J Hematol. 89(5): 656-63.
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