UCH-L1/PGP9.5 Monoclonal antibody

• Featured Product
• KD/KO Validated

UCHL1/PGP9.5 Monoclonal Antibody for WB, IHC, IF/ICC, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat, pig

Applications

WB, IHC, IF/ICC, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

11

CloneNo.

1C9E11

Cat no : 66230-1-Ig

Synonyms

PGP9.5, UCHL1, PGP 9.5, PARK5, Neuron cytoplasmic protein 9.5

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Validation Data Gallery

• 

  WB analysis using 66230-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66230-1-Ig (UCHL1 antibody) at dilution of 1:50000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated Beta Actin Monoclonal antibody (HRP-66009) as loading control.

• 

  WB analysis of Y79 using 66230-1-Ig

  Y79 cells were subjected to SDS PAGE followed by western blot with 66230-1-Ig (UCHL1 Antibody) at dilution of 1:50000 incubated at room temperature for 1.5 hours.

• 

  WB analysis of Neuro-2a using 66230-1-Ig

  Neuro-2a cells were subjected to SDS PAGE followed by western blot with 66230-1-Ig (UCHL1 Antibody) at dilution of 1:50000 incubated at room temperature for 1.5 hours.

• 

  IHC staining of mouse cerebellum using 66230-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue slide using 66230-1-Ig (UCHL1/PGP9.5 antibody) at dilution of 1:8000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat cerebellum using 66230-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue slide using 66230-1-Ig (UCHL1/PGP9.5 antibody) at dilution of 1:8000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat cerebellum using 66230-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue slide using 66230-1-Ig (UCHL1/PGP9.5 antibody) at dilution of 1:8000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse cerebellum using 66230-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue slide using 66230-1-Ig (UCHL1/PGP9.5 antibody) at dilution of 1:8000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of SH-SY5Y using 66230-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed SH-SY5Y cells using UCHL1/PGP9.5 antibody (66230-1-Ig, Clone: 1C9E11 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L), CL594-Phalloidin (red).

• 

  FC experiment of Y79 using 66230-1-Ig

  1x10^6 Y79 cells were intracellularly stained with 0.4 ug UCHL1/PGP9.5 Monoclonal antibody (66230-1-Ig, Clone:1C9E11) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) (SA00013-1)(red), or 0.4 ug Mouse IgG1 Isotype Control (MOPC-21) (65124-1-Ig, Clone: MOPC-21) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: Neuro-2a cells, Y79 cells, PC-12 cells, Pig brain tissue, Pig cerebellum tissue, Rat brain tissue, Rat cerebellum tissue, Mouse brain tissue, Mouse cerebellum tissue
• Positive IHC detected in: mouse cerebellum tissue, rat cerebellum tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF/ICC detected in: SH-SY5Y cells
• Positive FC (Intra) detected in: Y79 cells

Recommended dilution

• Western Blot (WB): WB : 1:20000-1:100000
• Immunohistochemistry (IHC): IHC : 1:4000-1:16000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• KD/KO: See 1 publications below
• WB: See 5 publications below
• IHC: See 3 publications below
• IF: See 4 publications below

Product Information

66230-1-Ig targets UCHL1/PGP9.5 in WB, IHC, IF/ICC, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat, pig samples.

• Tested Reactivity: human, mouse, rat, pig
• Cited Reactivity: human, mouse, rat, pig
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: UCHL1/PGP9.5 fusion protein Ag6547
• Full Name: ubiquitin carboxyl-terminal esterase L1 (ubiquitin thiolesterase)
• Calculated Molecular Weight: 25 kDa
• Observed Molecular Weight: 27 kDa
• GenBank Accession Number: BC000332
• Gene Symbol: UCHL1
• Gene ID (NCBI): 7345
• RRID: AB_2881621
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

Ubiquitin C-terminal hydrolase L1 (UCHL1) was originally identified as a neuronal protein that accounts for nearly 2% of total brain proteins. UCHL1 activity protects neurons from hypoxic injury, and binding of stroke-induced reactive lipid species to the cysteine 152 (C152) of UCHL1 unfolds the protein and disrupts its function. Reduced hydrolytic activity of mutant UCHL1 is implicated in the pathophysiologic process of Parkinson's and Alzheimer's disease due to abnormal neurotoxic protein aggregation. (PMID: 31356902, PMID: 30760601)

Publications

Species	Application	Title
human	WB	Theranostics; UCH-L1-mediated Down-regulation of Estrogen Receptor α Contributes to Insensitivity to Endocrine Therapy for Breast Cancer.; Authors - Xi-Sha Chen ; KD Validated
human	IHC	Sci Rep; Gingival proteomics reveals the role of TGF beta and YAP/TAZ signaling in Raine syndrome fibrosis; Authors - Cláudio Rodrigues Rezende Costa
rat	IHC	World J Gastroenterol; Hydrogen-rich water alleviates constipation by attenuating oxidative stress through the sirtuin1/nuclear factor-erythroid-2-related factor 2/heme oxygenase-1 signaling pathway; Authors - Kai-Di Chen
mouse	IF	Toxicol Appl Pharmacol; Ursolic acid alleviates paclitaxel-induced peripheral neuropathy through PPARγ activation; Authors - Yulian Yang
human	IF	Dig Dis Sci; P2Y1, P2Y2, and TRPV1 Receptors Are Increased in Diarrhea-Predominant Irritable Bowel Syndrome and P2Y2 Correlates with Abdominal Pain.; Authors - Yumei Luo
pig	WB, IF	Reprod Domest Anim; Isolation and in vitro expansion of porcine spermatogonial stem cells.; Authors - Xin Zhao

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