Fas/CD95 Cell Lysate | FAS cell lysate
Rat Fas/CD95 HEK293 Overexpression Lysate
Gene Names
Fas; Tnfrsf6
Applications
Western Blot
Synonyms
Fas/CD95; Rat Fas/CD95 HEK293 Overexpression Lysate; Rat FAS/CD95/APO-1/TNFRSF6 HEK293 Cell Lysate (WB positive control); Fas cell surface death receptor; FAS cell lysate
Host
HEK293 Cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading 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.
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.
Sequence Positions
390aa
Sequence
Met1-Lys170
Applicable Applications for FAS cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
Rat
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 200uL pure water and boil for 2-5 min.
Sequence Construction
A DNA sequence encoding the rat FAS (NP_631933.2) (Met1-Lys170) was expressed with the Fc region of human IgG1 at the C-terminus.
Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for FAS cell lysate
CD95 (APO-1/Fas) is an important inducer of the extrinsic apoptosis signaling pathway and therapy induced apoptosis of many tumor cells has been linked to the activity of CD95. is a prototype death receptor characterized by the presence of an 80 amino acid death domain in its cytoplasmic tail. This domain is essential for the recruitment of a number of signaling components upon activation by either agonistic anti-CD95 antibodies or cognate CD95 ligand that initiate apoptosis. The complex of proteins that forms upon triggering of CD95 is called the death-inducting signaling complex (DISC). The DISC consists of an adaptor protein and initiator caspases and is essential for induction of apoptosis. CD95 is also crucial for the negative selection of B cells within the germinal center (GC). Impairment of CD95-mediated apoptosis results in defective affinity maturation and the persistence of autoreactive B-cell clones. Changes in the expression of CD95 and/or its ligand CD95L are frequently found in human cancer. The downregulation or mutation of CD95 has been proposed as a mechanism by which cancer cells avoid destruction by the immune system through reduced apoptosis sensitivity. Thus, CD95 has therefore been viewed as a tumor suppressor. CD95 has been reported to be involved in the activation of NF-kappaB, MAPK3/ERK1, MAPK8/JNK, and the alternate pathways for CTL-mediated cytotoxicity. Accordingly, this protein is implicated in the pathogenesis of various malignancies and diseases of the immune system. The CD95/CD95L system was implicated in the etiology of inflammatory bowel disease (IBD) based, primarily, on the finding that CD95 is highly expressed in the intestinal epithelial cells and that epithelial apoptosis is increased in IBD.
This Rat Fas/CD95 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Fas/CD95 protein from the overexpression lysate was verified.
This Rat Fas/CD95 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Fas/CD95 protein from the overexpression lysate was verified.
Product Categories/Family for FAS cell lysate
References
Mschen M, et al. (2002) The origin of CD95-gene mutations in B-cell lymphoma. Trends Immunol. 23(2): 75-80.Peter ME, et al. (2003) The CD95(APO-1/Fas) DISC and beyond. Cell Death Differ. 10(1): 26-35.Peter ME, et al. (2005) Does CD95 have tumor promoting activities Biochim Biophys Acta. 1755(1): 25-36.Chen L, et al. (2010) Cell death in the colonic epithelium during inflammatory bowel diseases: CD95/Fas and beyond. Inflamm Bowel Dis. 16(6): 1071-6.
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
36,835 Da
NCBI Official Full Name
tumor necrosis factor receptor superfamily member 6
NCBI Official Synonym Full Names
Fas cell surface death receptor
NCBI Official Symbol
Fas
NCBI Official Synonym Symbols
Tnfrsf6
NCBI Protein Information
tumor necrosis factor receptor superfamily member 6; CD95 antigen; Fas receptor; apo-1 antigen; FASLG receptor; Fas antigen (ATP1); apoptosis-mediating surface antigen FAS; Fas (TNF receptor superfamily, member 6); Tumor necrosis factor receptor superfami
UniProt Protein Name
Tumor necrosis factor receptor superfamily member 6
Protein Family
UniProt Gene Name
Fas
UniProt Synonym Gene Names
Apt1; Tnfrsf6
UniProt Entry Name
TNR6_RAT
NCBI Description
Tnfsf6/Fasl receptor [RGD, Feb 2006]
Uniprot Description
Receptor for TNFSF6/FASLG. The adapter molecule FADD recruits caspase-8 to the activated receptor. The resulting death-inducing signaling complex (DISC) performs caspase-8 proteolytic activation which initiates the subsequent cascade of caspases (aspartate-specific cysteine proteases) mediating apoptosis. FAS-mediated apoptosis may have a role in the induction of peripheral tolerance, in the antigen-stimulated suicide of mature T-cells, or both ().
