Thimet Oligopeptidase 1 (TOP1)

Thimet oligopeptidase (THOP1), also known as endopeptidase EC 3.4.24.15 (EP24.15), is a zinc peptidase of the M3 family that also includes neurolysin/EC 3.4.24.16 and mitochondrial intermediate peptidase. Widely expressed by mammalian tissues and reported to be present in different subcellular locations, THOP1 is primarily a cytoplasmic enzyme capable of hydrolyzing a number of bioactive peptides and peptides released from by the proteosome, limiting antigenic presentation by MHC class I molecules. The amino acid sequence of human THOP1 is 90% and 86% identical to that of dog/mouse and rat.\n\nThimet oligopeptidase (TOP) is a thiol-dependent metallopeptidase, which can cleave and thereby modulate the activity of many neuropeptides. The enzyme is active in many endocrine tissues, including testis, brain, and pituitary. In rat, the richest source of TOP is the testes, with a specific activity fivefold that of brain. The mechanism whereby rat TOP expression is regulated at the transcriptional level has been examined by reporter gene assay and electromobility shift assays after isolation of 1020 bp of upstream sequence. Computer analysis predicts a number of potential transcription factor-binding sites, which were examined by deletion analysis and DNA-binding studies. The promoter or its deletion fragments were fused to luciferase reporter gene vectors and introduced into GH3 pituitary, COS-1 kidney, MAT-Lu prostate, and GC-2spd(ts) spermatid cells. Two regions of the promoter have been identified: a positively acting region (-901/-219) and a strong negatively acting region (-219/-102). Concomitantly, potential transcription factors interacting with the cis-acting elements of the promoter were studied by gel electromobility shift assays. This work has identified a number of transcription factor-binding sites. However, no differences in the binding behavior in the various cell lines was observed.\n\n\nApplications: \nWestern Blot: 0.1-0.2ug/ml with the appropriate secondary reagents to detect human THOP1. The detection limit for rhTHOP1 is approximately 5 ng/lane under non-reducing and reducing conditions. \nImmunoprecipitation: This antibody has been used to immunoprecipitate rhTHOP1 from conditioned media of transfected NS0 cells. \nDirect ELISA: This antibody can be used at 0.5-1.0ug/ml with the appropriate secondary reagents to detect human THOP1. The detection limit for rhTHOP1 is approximately 0.2 ng/well.\nOptimal dilutions should be determined by each laboratory for each application.\n\nStorage and Stability:\nLyophilized powder may be stored at 4 degrees C for short-term only. Reconstitute to nominal volume by adding sterile 40-50% glycerol and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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SPECIFICATIONS

Catalog Number

T5008-76

Size

100ug

Applications

ELISA, IP, WB

Hosts

Sheep

Reactivities

Hum

Form

Supplied as a lyophilized powder from a 0.2um filtered solution in PBS with 5% trehalose. Reconstitute with sterile PBS, 40-50% glycerol. If 0.5ml is used, the concentration will be 0.2mg/ml.

P Type

Pab

Purity

Purified by human THOP1 affinity chromatography.

Isotype

IgG

References

General References:\n1. Conn, K. J., et al., J. Neurochem. 66: 2011-2018, 1996.\n2. Yamin, R., et al., J. Biol. Chem. 274: 18777-19784, 1999.\n3. York, I. A., et al., Immunity 18: 429-440, 2003.\n4. Suzan McCool, Adrian R. Pierotti. DNA and Cell Biology. December 1, 2000, 19(12): 729-738. doi:10.1089

Additional Info

Selected for its ability to recognize human THOP1 in direct ELISAs and Western Blots. In Western Blots, this antibody shows less than 5% crossreactivity with rhNeurolysin.