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PGP9.5 Mouse Monoclonal Antibody [B1-5-6]

Usd: 350

Specification

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Catalog# EM1701-86

PGP9.5 Mouse Monoclonal Antibody [B1-5-6]

Application

  • WB

  • IHC-P

  • IF-Cell

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

PGP9.5 Mouse Monoclonal Antibody [B1-5-6]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Synthetic peptide within Human PGP95 aa 174-223 / 223.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IHC-P, IF-Cell, FC

Molecular Weight

Predicted band size: 25 kDa

Positive Control

A-172 cell lysate, SHG-44 cell lysate, U-87 MG cell lysate, SH-SY5Y cell lysate, NCI-H1299 cell lysate, A549 cell lysate, 293T cell lysate, Neuro-2a cell lysate, C6 cell lysate, PC-12 cell lysate, mouse brain tissue lysate, rat brain tissue lysate, SH-SY-5Y, rat cerebellum tissue, human pancreas tissue, human kidney tissue, mouse brain tissue, Neuro-2a, PC-12.

Conjugation

unconjugated

Clone Number

B1-5-6

RRID

AB_3068809

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG2a

Purification Method

Immunogen affinity purified.

Application Dilution

  • WB

  • 1:2,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:50-1:200

  • FC

  • 1:1,000

Target

Function

Ubiquitin-protein hydrolase involved both in the processing of ubiquitin precursors and of ubiquitinated proteins. This enzyme is a thiol protease that recognizes and hydrolyzes a peptide bond at the C-terminal glycine of ubiquitin. Also binds to free monoubiquitin and may prevent its degradation in lysosomes. The homodimer may have ATP-independent ubiquitin ligase activity.

Background References

1. Wada H et al. Cleavage of the C-terminus of NEDD8 by UCH-L3. Biochem Biophys Res Commun 251:688-692 (1998).

2. Liu Y et al. The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinson\'s disease susceptibility. Cell 111:209-218 (2002).

Sequence Similarity

Belongs to the peptidase C12 family.

Tissue Specificity

Found in neuronal cell bodies and processes throughout the neocortex (at protein level). Expressed in neurons and cells of the diffuse neuroendocrine system and their tumors. Weakly expressed in ovary. Down-regulated in brains from Parkinson disease and Alzheimer disease patients.

Post-translational Modification

O-glycosylated.

Subcellular Location

Cytoplasm. Endoplasmic reticulum.

UNIPROT

SwissProt: P09936 Human

SwissProt: Q9R0P9 Mouse

SwissProt: Q00981 Rat

Synonyms

Epididymis luminal protein 117 antibody

Epididymis secretory protein Li 53 antibody

HEL 117 antibody

HEL S 53 antibody

NDGOA antibody

Neuron cytoplasmic protein 9.5 antibody

OTTHUMP00000218137 antibody

OTTHUMP00000218139 antibody

OTTHUMP00000218140 antibody

OTTHUMP00000218141 antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Relative expression (RE)</span><br /><br />Western blot analysis of PGP9.5 on different lysates with Mouse anti-PGP9.5 antibody (<a href="/products/EM1701-86" style="font-weight: bold;text-decoration: underline;">EM1701-86</a>) at 1/2,000 dilution.<br /><br />Lane 1: A-172 cell lysate (20 µg/Lane)<br />Lane 2: SHG-44 cell lysate (20 µg/Lane)<br />Lane 3: U-87 MG cell lysate (20 µg/Lane)<br />Lane 4: SH-SY5Y cell lysate (20 µg/Lane)<br />Lane 5: NCI-H1299 cell lysate (20 µg/Lane)<br />Lane 6: A549 cell lysate (20 µg/Lane)<br />Lane 7: 293T cell lysate (20 µg/Lane)<br />Lane 8: LNCaP cell lysate (negative) (20 µg/Lane)<br />Lane 9: K-562 cell lysate (negative) (20 µg/Lane)<br />Lane 10: Neuro-2a cell lysate (20 µg/Lane)<br />Lane 11: C6 cell lysate (20 µg/Lane)<br />Lane 12: PC-12 cell lysate (20 µg/Lane)<br />Lane 13: Mouse brain tissue lysate (20 µg/Lane)<br />Lane 14: Rat brain tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 25 kDa<br />Observed band size: 25 kDa<br /><br />Exposure time: 1 minute 50 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/EM1701-86" style="font-weight: bold;text-decoration: underline;">EM1701-86</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Relative expression (RE)

    Western blot analysis of PGP9.5 on different lysates with Mouse anti-PGP9.5 antibody (EM1701-86) at 1/2,000 dilution.

    Lane 1: A-172 cell lysate (20 µg/Lane)
    Lane 2: SHG-44 cell lysate (20 µg/Lane)
    Lane 3: U-87 MG cell lysate (20 µg/Lane)
    Lane 4: SH-SY5Y cell lysate (20 µg/Lane)
    Lane 5: NCI-H1299 cell lysate (20 µg/Lane)
    Lane 6: A549 cell lysate (20 µg/Lane)
    Lane 7: 293T cell lysate (20 µg/Lane)
    Lane 8: LNCaP cell lysate (negative) (20 µg/Lane)
    Lane 9: K-562 cell lysate (negative) (20 µg/Lane)
    Lane 10: Neuro-2a cell lysate (20 µg/Lane)
    Lane 11: C6 cell lysate (20 µg/Lane)
    Lane 12: PC-12 cell lysate (20 µg/Lane)
    Lane 13: Mouse brain tissue lysate (20 µg/Lane)
    Lane 14: Rat brain tissue lysate (20 µg/Lane)

    Predicted band size: 25 kDa
    Observed band size: 25 kDa

    Exposure time: 1 minute 50 seconds; ECL: K1801;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1701-86) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Knockdown (KD)</span><br /><br />Western blot analysis of PGP9.5 on different lysates with Mouse anti-PGP9.5 antibody (<a href="/products/EM1701-86" style="font-weight: bold;text-decoration: underline;">EM1701-86</a>) at 1/2,000 dilution.<br /><br />Lane 1: A549-si NT cell lysate (10 µg/Lane)<br />Lane 2: A549-si PGP9.5 cell lysate (10 µg/Lane)<br /><br />Predicted band size: 25 kDa<br />Observed band size: 25 kDa<br /><br />Exposure time: 30 seconds; ECL: K1802;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/EM1701-86" style="font-weight: bold;text-decoration: underline;">EM1701-86</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Knockdown (KD)

    Western blot analysis of PGP9.5 on different lysates with Mouse anti-PGP9.5 antibody (EM1701-86) at 1/2,000 dilution.

    Lane 1: A549-si NT cell lysate (10 µg/Lane)
    Lane 2: A549-si PGP9.5 cell lysate (10 µg/Lane)

    Predicted band size: 25 kDa
    Observed band size: 25 kDa

    Exposure time: 30 seconds; ECL: K1802;
    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (EM1701-86) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of Neuro-2a cells labeling PGP9.5 with Mouse anti-PGP9.5 antibody (<a href="/products/EM1701-86" style="font-weight: bold;text-decoration: underline;">EM1701-86</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-PGP9.5 antibody (<a href="/products/EM1701-86" style="font-weight: bold;text-decoration: underline;">EM1701-86</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of Neuro-2a cells labeling PGP9.5 with Mouse anti-PGP9.5 antibody (EM1701-86) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-PGP9.5 antibody (EM1701-86) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.

  • Immunocytochemistry analysis of PC-12 cells labeling PGP9.5 with Mouse anti-PGP9.5 antibody (<a href="/products/EM1701-86" style="font-weight: bold;text-decoration: underline;">EM1701-86</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-PGP9.5 antibody (<a href="/products/EM1701-86" style="font-weight: bold;text-decoration: underline;">EM1701-86</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />beta Tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of PC-12 cells labeling PGP9.5 with Mouse anti-PGP9.5 antibody (EM1701-86) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Mouse anti-PGP9.5 antibody (EM1701-86) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human pancreas tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.

  • Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.

    Immunohistochemical analysis of paraffin-embedded mouse brain tissue using anti-PGP9.5 antibody. Counter stained with hematoxylin.

  • Flow cytometric analysis of Neuro-2a cells labeling PGP9.5.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/EM1701-86" style="font-weight: bold;text-decoration: underline;">EM1701-86</a>, 1μg/mL) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Mouse IgG Secondary antibody (<a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of Neuro-2a cells labeling PGP9.5.

    Cells were fixed and permeabilized. Then stained with the primary antibody (EM1701-86, 1μg/mL) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

  • Flow cytometric analysis of PC-12 cells labeling PGP9.5.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/EM1701-86" style="font-weight: bold;text-decoration: underline;">EM1701-86</a>, 1μg/mL) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor&trade; 488 conjugate-Goat anti-Mouse IgG Secondary antibody (<a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    Flow cytometric analysis of PC-12 cells labeling PGP9.5.

    Cells were fixed and permeabilized. Then stained with the primary antibody (EM1701-86, 1μg/mL) (red) compared with Mouse IgG1 Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Citation

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