Human/Mouse/Rat UCH-L1/PGP9.5 Antibody

Catalog #: MAB60072 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB60072-100
MAB60072-SP

Detection of Human, Mouse, and Rat UCH-L1/PGP9.5 by Western Blot.

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Human/Mouse/Rat UCH-L1/PGP9.5 Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects human UCH-L1/PGP9.5 in direct ELISAs. Detects human, mouse, and rat in Western blot.

Source

Monoclonal Mouse IgG_2B Clone # 972119

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant human UCH-L1/PGP9.5
Gln2-Ala223
Accession # P09936

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

See below

Simple Western

1 µg/mL

See below

Immunohistochemistry

5-25 µg/mL

See below

Immunocytochemistry

0.3-25 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human, Mouse, and Rat UCH-L1/PGP9.5 by Western Blot. Western blot shows lysates of A172 human glioblastoma cell line, Neuro-2A mouse neuroblastoma cell line, and PC-12 rat adrenal pheochromocytoma cell line. PVDF membrane was probed with 0.1 µg/mL of Mouse Anti-Human UCH-L1/PGP9.5 Monoclonal Antibody (Catalog # MAB60072) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for UCH-L1/PGP9.5 at approximately 26 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

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UCH-L1/PGP9.5 in A172 Human Cell Line. UCH-L1/PGP9.5 was detected in immersion fixed A172 human glioblastoma cell line using Mouse Anti-Human UCH-L1/PGP9.5 Monoclonal Antibody (Catalog # MAB60072) at 0.3 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Immunohistochemistry

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UCH-L1/PGP9.5 in Human Brain. UCH-L1/PGP9.5 was detected in immersion fixed paraffin-embedded sections of human brain (caudate nucleus) using Mouse Anti-Human UCH-L1/PGP9.5 Monoclonal Antibody (Catalog # MAB60072) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Simple Western

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Detection of Human and Mouse UCH-L1/PGP9.5 by Simple Western^TM. Simple Western lane view shows lysates of A172 human glioblastoma cell line and Neuro‑2A mouse neuroblastoma cell line, loaded at 0.2 mg/mL. A specific band was detected for UCH-L1/PGP9.5 at approximately 31 kDa (as indicated) using 1 µg/mL of Mouse Anti-Human UCH-L1/PGP9.5 Monoclonal Antibody (Catalog # MAB60072). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: UCH-L1/PGP9.5

UCH-L1 (ubiquitin carboxyterminal hydrolase isozyme 1; also PGP9.5) is a 24-27 kDa member of the peptidase C12 family of enzymes. It shows restricted expression, being found in neurons and oocytes. UCH-L1 has dual enzymatic activity. As a monomer, it is a ubiquitin hydrolase that removes ubiquitin from modified proteins; as a homodimer, it acts as a ligase that creates ubiquitin dimers. In neurons, UCH-L1’s most important role appears to be that of generating free ubiquitin. Human UCH-L1 is 223 amino acids (aa) in length. It is O-glycosylated, ubiquitinated, and farnesylated; when farnesylated, it becomes associated with cell membranes. Three potential splice forms are reported. One shows a two aa substitution for aa 12-15, a second contains an alternative start site at Met82, and a third shows the same start site coupled with a deletion of aa 138-153. Full-length human UCH-L1 shares 95% aa identity with mouse UCH-L1.

Long Name

Ubiquitin C-terminal Hydrolase L1

Entrez Gene IDs

7345 (Human); 22223 (Mouse)

Alternate Names

EC 3.4.19.12; EC 6.-; Neuron cytoplasmic protein 9.5; PARK5; PGP 9.5; PGP9.5; PGP9.5Uch-L1; PGP95; ubiquitin carboxyl-terminal esterase L1 (ubiquitin thiolesterase); ubiquitin carboxyl-terminal hydrolase isozyme L1; ubiquitin C-terminal hydrolase; Ubiquitin thioesterase L1; UCHL1; UCH-L1