UCH-L1/PGP9.5 Antibody

Images

 
Western Blot: PGP9.5 / UCHL-1 Antibody [NB300-675] - Western blot analysis of extracts from C6 cells using PGP9.5 / UCHL-1 antibody (NB300-675, 1:200). Image from verified customer review.
Immunocytochemistry/ Immunofluorescence: PGP9.5 / UCHL-1 Antibody [NB300-675] - IF Confocal analysis of C6 cells using PGP9.5 / UCHL-1 antibody (NB300-675, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used ...read more
Immunohistochemistry: PGP9.5 / UCHL-1 Antibody [NB300-675] - Human brain, Substantia Nigra, Pigmented Neuron, 40X.
Western Blot: PGP9.5 / UCHL-1 Antibody [NB300-675] - Detection of UCHL1 in mouse brain lysate.
Immunocytochemistry/ Immunofluorescence: PGP9.5 / UCHL-1 Antibody [NB300-675] - in Neuro2a cells with FITC (green). Nuclei were counterstained with Dapi (blue).
Immunofluorescence: PGP9.5 / UCHL-1 Antibody [NB300-675] - analysis of acetone-fixed, frozen mouse ear skin section using anti-PGP9.5 antibody. Image from verified customer review.

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Bv, Eq, GP, PmSpecies Glossary
Applications WB, ICC/IF, IHC, IHC-P
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1 mg/ml

UCH-L1/PGP9.5 Antibody Summary

Immunogen
A synthetic peptide made to an internal portion of the human UCHL1 protein (between residues 100-200). [UniProt# P09936]
Localization
Cytoplasmic
Marker
pan-Neuronal Marker
Predicted Species
Rat (99%), Porcine (99%), Bovine (92%), Guinea Pig (100%), Primate (100%), Equine (99%). Backed by our 100% Guarantee.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
UCHL1
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 0.5-2.0 ug/ml
  • Immunocytochemistry/Immunofluorescence 1:100-1:200
  • Immunohistochemistry 2.5-5.0 ug/ml
  • Immunohistochemistry-Paraffin 2.5-5.0 ug/ml
Application Notes
This UCHL1 antibody is useful for Immunocytochemistry/Immunofluorescence, Western blot and Immunohistochemistry-Paraffin. By Western blot a band at ~25 kDa is seen.

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.
Theoretical MW
25 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.
Positive Control
Brain Lysate (NB820-59657)
UCH-L1/PGP9.5 Lysate (NBL1-17575)
Reviewed Applications
Read 5 Reviews rated 5
using
NB300-675 in the following applications:

Reactivity Notes

Human and mouse. Predicted to react with primate and guinea pig based on 100% sequence homology. Immunogen sequence has 99% homology to rat, pig, and horse, and 92% homology to bovine.

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
Tris-Glycine and 0.15M NaCl
Preservative
0.05% Sodium Azide
Concentration
1 mg/ml
Purity
Immunogen affinity purified

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.

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Applications: WB, ICC/IF, IHC, IHC-P

Publications for UCH-L1/PGP9.5 Antibody (NB300-675) (0)

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Reviews for UCH-L1/PGP9.5 Antibody (NB300-675) (5) 55

Average Rating: 5
(Based on 5 reviews)
We have 5 reviews tested in 4 species: Human, Mouse, Rat, opossum.
We have 5 reviews tested in 4 application: IHC-Fr, IF, WB, ICC.

Reviews using NB300-675:
Filter by Applications
IHC-Fr
(2)
IF
(1)
WB
(1)
ICC
(1)
All Applications
Filter by Species
Human
(2)
Mouse
(1)
Rat
(1)
opossum
(1)
All Species
Images Ratings Applications Species Date Details
Immunohistochemistry-Frozen UCH-L1/PGP9.5 NB300-675
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reviewed by:
Anonymous
IHC-Fr opossum 01/04/2017
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Summary

ApplicationImmunohistochemistry-Frozen
Sample TestedTestis
Speciesopossum

Comments

CommentsTested in gray short-tailed opossum (Monodelphis domestica) in testis from 8-month animal.
Tissue was fixed in 4% Paraformaldehyde overnight, incubated in 30% sucrose/PBS overnight and embedded in OCT. Sections were cut on a cryostat at 10um.

For immunofluorescence staining: Slides were dried 15 minutes, washed 3x3 min TBST (0.1% Triton X-100), permeabilized for 10 minutes in 2% Triton X-100 in TBS, rinsed 3x3 min TBST, blocked in 10% heat inactivated goat serum in TBST for 30 min, and primary added at 1:100 in 10% blocking solution. Slides were coverslipped with parafilm overnight at 4C. The next day slides were washed with TBST 4x5 min, secondaries (AlexaFluor 568 goat anti rabbit and AlexaFluor 488 goat anti mouse 1:500) for 1 hour, 3x3 washes TBST, and mounted with VectaShield HardSet (+ DAPI).

Intense staining at periphery of seminiferous tubules of Uchl-1 (red in image) with these conditions. 1:200 also worked though staining was less intense (not shown). The same conditions with 1% Triton X-100 for permeabiliziation resulted in weak staining at 1:100 and 1:200. However, ID-4 did not work well under any conditions. Shown in green is some faint staining in interstitial cells but this was not consistent between samples and did not appear to be specific. ID-4 was tested in 4 and 8 month opossum testes, and at dilutions ranging from 1:25 to 1:500. Single-antibody staining was also performed for each antibody and successful in Uchl-1, not ID-4.
 
reviewed by:
Anonymous
IHC-Fr Rat 09/15/2015
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Summary

ApplicationImmunohistochemistry-Frozen
Sample TestedRat brain
SpeciesRat
Immunofluorescence UCH-L1/PGP9.5 NB300-675
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reviewed by:
Anonymous A
IF Mouse 08/29/2015
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Summary

ApplicationImmunofluorescence
Sample TestedMouse Ear Skin (frozen section)
SpeciesMouse

Blocking

Blocking Details1xPBS+ 3%BSA+5% goat serum, room temperature 1hour

Primary Anitbody

Dilution Ratio1:200 dilution, 4C, overnight

Secondary Antibody

Secondary Descriptiongoat anti-rabbit IgG, HRP
Secondary Concentration2.5ug/ml (1:200)

Details

Detection NotesZeiss microscope detection, rabbit IgG istotype control
Fixation Detailsacetone 5min fixation
Wash Description1xPBS imerse 3times, 5min each
Western Blot UCH-L1/PGP9.5 NB300-675
Enlarge
reviewed by:
Anony- mous
WB Human 07/01/2014
View

Summary

ApplicationWestern Blot
SpeciesHuman
Special ApplicationsWestern blot analysis of extracts from C6 cells using PGP9.5 / UCHL-1 antibody (NB300-675, 1:200).
FileView PDF
Immunocytochemistry UCH-L1/PGP9.5 NB300-675
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reviewed by:
Anony- mous
ICC Human 07/01/2014
View

Summary

ApplicationImmunocytochemistry
SpeciesHuman
Special ApplicationsIF Confocal analysis of C6 cells using PGP9.5 / UCHL-1 antibody (NB300-675, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green). Actin filaments were labeled with Alexa Fluor 568 phalloidin (red). DAPI was used to stain the cell nuclei (blue).
FileView PDF

Product General Protocols

View specific protocols for UCH-L1/PGP9.5 Antibody (NB300-675): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for UCH-L1/PGP9.5 Antibody (NB300-675) (0)

There are no specific FAQs related to this product. Read our general customer & technical service FAQs.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB300-675AF405
Alexa Fluor 488 Labeled NB300-675AF488
Alexa Fluor 647 Labeled NB300-675AF647
Alexa Fluor 700 Labeled NB300-675AF700
Biotin Labeled NB300-675B
DyLight 405 Labeled NB300-675V
DyLight 488 Labeled NB300-675G
DyLight 550 Labeled NB300-675R
DyLight 650 Labeled NB300-675C
FITC Labeled NB300-675F
HRP Labeled NB300-675H

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Recent Reviews

5
5
5
4
0
3
0
2
0
1
0

Anonymous
01/04/2017
Application: IHC-Fr
Species: opossum

 
Anonymous
09/15/2015
Application: IHC-Fr
Species: Rat

Anonymous A
08/29/2015
Application: IF
Species: Mouse

Bioinformatics

Gene Symbol UCHL1
Entrez