This page introduces methods for detecting apoptosis using Fas Ligand assay and links to relevant resources and papers.
In general, we have been looking at cell death in cultured adherant neuronal cells using confocal and laser scanning microscopy. We have initially used the TUNEL and Annexin techniques, there is a huge variety of techniques/analysis and applications used by different Labs.
Note also that many links on this page are to external internet resources.
OMIM - TUMOR NECROSIS FACTOR LIGAND SUPERFAMILY, MEMBER 6; TNFSF6 (FAS LIGAND; FASL APOPTOSIS ANTIGEN LIGAND 1; APT1LG1 APOPTOSIS ANTIGEN LIGAND CD95 LIGAND; CD95L CD178 ANTIGEN; CD178)
Emdbiosciences QIA27 | PDF QIA27 user protocol
CD95L ELISA Kit
Format: 96-well plate
Form: 96 Tests
Detection Method: Colorimetric
Species Reactivity: human
Specificity: Human Fas ligand (membrane bound and soluble)
Sensitivity: 0.02 ng/ml
Assay Range: 325 pg/ml
Assay Time: 4 h
Sample Type: Cell lysates, tissue culture medium, serum, and plasma
Kit Contents: Coated 96-well plate, FasL standard, detector antibody, 400X conjugate, conjugate diluent, substrate, sample diluent, wash concentrate, antigen extract agent, stop solution, plate sealers, and a user protocol. Comments: Suitable for the in vitro quantitation of human FasL. Recognizes both membrane bound and soluble FasL.
Towfighi A, Skolasky RL, St Hillaire C, Conant K, McArthur JC. CSF soluble Fas correlates with the severity of HIV-associated dementia. Neurology. 2004 Feb 24;62(4):654-6. Review.
Sabri F, De Milito A, Pirskanen R, Elovaara I, Hagberg L, Cinque P, Price R, Chiodi F. Elevated levels of soluble Fas and Fas ligand in cerebrospinal fluid of patients with AIDS dementia complex. J Neuroimmunol. 2001 Mar 1;114(1-2):197-206.
"Measurable levels of sFasL were found in 79% of the CSF and 98% of sera samples. ...CSF sFas (median and IQR 116, 132 vs. 30, 23 pg/ml, P less than 0.001) and sFasL (median and IQR 127, 290 vs. 15, 73 pg/ml, P less than 0.001) levels. The sFas in serum differed significantly between HIV-1 infected subjects and non-infected controls (Pless than 0.001), with no correlation to ADC. On the contrary, sFasL in serum differed among HIV-1 infected subjects according to clinical signs of ADC."
Felderhoff-Mueser U, Herold R, Hochhaus F, Koehne P, Ring-Mrozik E, Obladen M, Buhrer C. Increased cerebrospinal fluid concentrations of soluble Fas (CD95/Apo-1) in hydrocephalus. Arch Dis Child. 2001 Apr;84(4):369-72.
"High concentrations of sFas were observed in children with hydrocephalus (median 252 ng/ml); in controls sFas was below the detection limit (0.5 ng/ml). sFasL was undetectable in all but one sample."
Demjen D, Klussmann S, Kleber S, Zuliani C, Stieltjes B, Metzger C, Hirt UA, Walczak H, Falk W, Essig M, Edler L, Krammer PH, Martin-Villalba A. Neutralization of CD95 ligand promotes regeneration and functional recovery after spinal cord injury. Nat Med. 2004 Apr;10(4):389-95.
There are many different methods for determining programmed cell death. This page covers only those intended for use in our Lab.
Please email Dr Mark Hill if you wish to make a comment about this current project.