* Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own experiment using appropriate negative and positive controls.
Ammonium sulfate precipitation
10mM sodium azide
Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
Product Specific Information
Suggested positive control: antigen standard for UCHL1 (transient overexpression lysate), rat spinal cord or perpheral nerve homogenate.
Ubiquitin C-terminal hydrolase 1 (UCHL1) was originally identified as a major component of the neuronal cytoplasm from 2-dimensional gel analysis of brain tissues, and was given the name PGP9.5. The protein is extremely abundant representing 1-2% of total brain protein. It was later found that a ubiquitin C-terminal hydrolase enzyme activity was associated with the PGP9.5 protein, resulting in the renaming of PGP9.5 to ubiquitin C-terminal hydrolase 1. This is the first of a family of ubiquitin C-terminal hydrolases which have been characterized, and is expressed heavily in neurons in the brain. The ubiquitin C-terminal hydrolases cleave ubiquitin from other molecules. This activity is important to generate mono-ubiquitin from genes which encode polyubiquitin chains or ubiquitin fused to other proteins. The activity is also important to remove ubiquitin from partially degraded proteins, allowing the ubiquitin monomer to be recycled. Regulation of the ubiquitin pathway is very important and many disease states are associated with defects in this pathway. Point mutations in the UCHL1 gene are associated with some forms of human Parkinson's disease. Recent studies suggest that UCHL1 also has a ubiqutinyl ligase activity, being able to couple ubiquitin monomers by linking the C-terminus of one with lysine 63 of the other. Since UCHL1 is heavily expressed in neurons, antibodies to UCHL1 can be used to identify neurons in histological sections and in tissue culture. The great abundance of this protein in neurons means that it is released from neurons in large amounts following injury or degeneration, so the detection of of UCHL1 in CSF and other bodily fluids can be used as a biomarker. UCHL1 was also discovered as a gene mutated in some rare famial forms of Parkinson's disease. Interestingly a common allelic variant of UCHL1, the S18Y polymorphism is actually protective against Parkinson's disease.
UCHL1 Antibody (PA1-46204) in WB PA1-46204_Westernblot.jpg
Western Blot with anti-UCHL1 Polyclonal Antibody (PA1-46204)
Western blot of whole cell homogenate of the human SH-SY5Y neuroblastoma cell line stained with chicken antibody to glyceraldehyde 3 phosphate dehydrogenase (GAPDH, left lane, blot made with monoclonal antibody to GAPDH) and PA1-46204 (right lane). The GAPDH antibody stains a ~37kDa band while the UCHL1 antibody stains a band at about 24kDa.
UCHL1 Antibody (PA1-46204) in IF PA1-46204_Immunohistochemistry.jpg
Immunofluorescence with anti-UCHL1 Polyclonal Antibody (PA1-46204)
Shows rat mixed neuron/glial cultures stained with chicken UCHL1 (green) and rabbit antibody to glial fibrillary acidic protein (GFAP-red); Blue is a DNA stain. Note that the UCHL1 stains neurons strongly and specifically, and that the staining is concentrated in the cell bodies, though some does extend into the dendrites also.
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