UCH-L1/PGP9.5 Antibody

Western Blot: UCH-L1/PGP9.5 Antibody [NB110-58874] - Analysis of different tissue and cell lysates using rabbit pAb to UCHL1, NB110-58874, dilution 1:2,000 in red, and mouse mAb to HSP60, dilution 1:10,000, in green: [1] protein standard, [2] rat brain, [3] mouse brain, [4] NIH-3T3, [5] HEK293, [6] HeLa, [7] SH-SY5Y cells. The single band at 24kDa corresponds to the UCHL1 protein, while the 60kDa band represents HSP60 protein. UCHL1 is detectable in CNS extracts and cells with neuronal properties but not in HeLa, NIH-3T3 and other non-neuronal cells.

Immunocytochemistry/Immunofluorescence: UCH-L1/PGP9.5 Antibody [NB110-58874] - Analysis of SH-SY5Y cells stained with rabbit pAb to UCHL1, NB110-58874, dilution 1:1,000 in green, and costained with mouse mAb to fibrillarin, dilution 1:1,000 in red. Blue is DAPI staining of nuclear DNA. The UCHL1 antibody produces strong staining of the cellular cytoplasm of these cells which share many properties with neurons, while the fibrilarin antibody specifically labels nucleoli.

Western Blot: PGP9.5 / UCHL-1 Antibody [NB110-58874] - Blot of whole bovine brain extract stained with RPCA-UCHL1 showing a strong and clean band at about 24kDa.

Immunocytochemistry/Immunofluorescence: PGP9.5 / UCHL-1 Antibody [NB110-58874] - Shows rat mixed neuron/glial cultures stained with Rabbit anti-UCHL1 antibody (green). 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.

Simple Western: PGP9.5 / UCHL-1 Antibody [NB110-58874] - Simple Western lane view shows a specific band for PGP9.5/UCHL-1 in 0.05 mg/ml of Human Brain lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

• Reactivity: Hu, Mu, Rt, Po, Bv, Ch, Eq
• Applications: WB, Simple Western, ICC/IF, IHC
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated

UCH-L1/PGP9.5 Antibody Summary

• Description: Novus Biologicals Rabbit UCH-L1/PGP9.5 Antibody (NB110-58874) is a polyclonal antibody validated for use in IHC, WB, ICC/IF and Simple Western. Anti-UCH-L1/PGP9.5 Antibody: Cited in 2 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.
• Immunogen: Recombinant full length human UCHL1 purified from E. coli. [UniProt# P09936]
• Marker: pan-Neuronal Marker
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: UCHL1
• Purity: Unpurified

Applications/Dilutions

• Dilutions:
  • Immunocytochemistry/ Immunofluorescence 1:500
  • Immunohistochemistry 1:500
  • Immunohistochemistry-Paraffin
  • Simple Western 1:2000
  • Western Blot 1:5000
• Application Notes: This UCHL1 antibody is useful for Immunocytochemistry/Immunofluorescence, ABC or other enzyme linked immunocytochemical procedures and Western blotting. Use in IHC-P reported in scientific literature (PMID: 22527779).
  
  In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
  See Simple Western Antibody Database for Simple Western validation: Tested in Human Brain lysate 0.05 mg/mL, separated by Size, antibody dilution of 1:2000, apparent MW was 29 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.
• Theoretical MW: 24 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  UCH-L1/PGP9.5 Overexpression Lysate

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: Supplied as serum
• Preservative: 0.035% Sodium Azide
• Purity: Unpurified

Alternate Names for UCH-L1/PGP9.5 Antibody

• 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

Background

UCH-L1 (ubiquitin carboxyl-terminal hydrolase isozyme L1) was the first discovered de-ubiquitinating enzyme which implicates in processing of ubiquitin precursors and also of ubiquitinated proteins. UCH-L1 is a thiol protease that recognizes and hydrolyzes a peptide bond at C-terminal glycine of ubiquitin. It also binds to free monoubiquitin and prevents its degradation in lysosomes. Localized in cytoplasm and ER membrane as lipid-anchor, UCH-L1 expression is restricted to brain, peripheral nerves, endocrine tissues and gonads of both sexes etc. UCH-L1 deletion in mice leads to fatal neurodegenerative disorder known as gracile axonal dystrophy and it is down-regulated in brains from Parkinson as well as Alzheimer disease patients. Expression outside of neuro-endocrine tissues is found in various cancers including B-cell lymphoma, multiple myeloma, and lung cancer. In transgenic mouse model, UCH-L1 has been demonstrated as an oncogene that causes malignancies by boosting AKT signalling. Furthermore, UCH-L1 has been shown to interfere with ubiquitination of RAPTOR which is catalyzed by DDB1-Cul4 E3 ligase complex, leading to loss of mTORC1 integrity accompanied by a concurrent increase in mTORC2, likely due to increased availability of free mTOR.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for UCH-L1/PGP9.5 Antibody (NB110-58874)(2)

Publications using NB110-58874

• Vincent K, Mohanty S, Pinelli R et al. Aging of mouse intervertebral disc and association with back pain Bone 2019-03-29 [PMID: 30936040] (ICC/IF, Mouse)
• Marmotti A, Cravino M, Germano M et al. Peroneal tendoscopy. Curr Rev Musculoskelet Med. 2012-06-01 [PMID: 22527779] (IHC-P, Human)

Bioinformatics

• Gene Symbol: UCHL1
• Entrez:
  • 7345(Human)
  • 29545(Rat)
• Uniprot:
  • P09936(Human)
  • Q00981(Rat)