使用敲除细胞株进行验证

Anti-PGP9.5抗体[346CT2.5.1] (ab86808)

概述

  • 产品名称
    Anti-PGP9.5抗体[346CT2.5.1]
    参阅全部 PGP9.5 一抗
  • 描述
    小鼠单克隆抗体[346CT2.5.1] to PGP9.5
  • 经测试应用
    适用于: IHC-P, Flow Cyt, WBmore details
  • 种属反应性
    与反应: Rat, Goat, Cat, Dog, Human
    预测可用于: Mouse, Horse, Pig
  • 免疫原

    Recombinant full length protein

  • 阳性对照
    • This antibody gave a positive signal in the following lysates: Human Brain Tissue; Rat Brain Tissue; Rat Brain Cortex Tissue; PC12 Whole Cell; Y79 Whole Cell; U-87MG Whole Cell; SHSY-5Y Whole Cell and HAP1 Wole cells.
  • 常规说明

    This antibody clone is manufactured by Abcam.

    This monoclonal antibody to PGP9.5 has been knockout validated in Western blot. The expected band for PGP9.5 was observed in wild type cells and the band was not seen in knockout cells.

性能

应用

Our Abpromise guarantee covers the use of ab86808 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
IHC-P 1/1000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt Use 0.01-0.1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).

Abcam recommends blocking with 3% Milk.

靶标

  • 功能
    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.
  • 组织特异性
    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.
  • 疾病相关
    Parkinson disease 5
    Neurodegeneration with optic atrophy, childhood-onset
  • 序列相似性
    Belongs to the peptidase C12 family.
  • 翻译后修饰
    O-glycosylated.
  • 细胞定位
    Cytoplasm. Endoplasmic reticulum membrane. About 30% of total UCHL1 is associated with membranes in brain.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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
    • Park 5 antibody
    • PARK5 antibody
    • PGP 9.5 antibody
    • PGP9.5 antibody
    • PGP95 antibody
    • Protein gene product 9.5 antibody
    • Ubiquitin C terminal esterase L1 antibody
    • Ubiquitin C terminal hydrolase antibody
    • Ubiquitin C terminal hydrolase L1 antibody
    • Ubiquitin carboxyl terminal esterase L1 antibody
    • Ubiquitin carboxyl terminal hydrolase isozyme L1 antibody
    • Ubiquitin carboxyl-terminal hydrolase isozyme L1 antibody
    • Ubiquitin thioesterase L1 antibody
    • Ubiquitin thiolesterase antibody
    • Ubiquitin thiolesterase L1 antibody
    • UCH-L1 antibody
    • UCHL1 antibody
    • UCHL1_HUMAN antibody
    see all

图片

  • IHC-P image of PGP9.5 staining on cat bladder sections using ab86808 (1:3000). The sections were subjected to heat mediated antigen retrieval using citric acid. The sections were then blocked using 1% BSA at 21°C for 10 min. ab86808 was incubated at 21°C for 1 hours. The secondary antibody was Goat polyclonal to anti-mouse (rat pre-absorbed) conjugated to biotin (1:200).

    See Abreview



  • Predicted band size : 25 kDa

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: UCHL1 (KO) knockout  HAP1 whole cell lysate (20 µg)
    Lane 3: SH-SY5Y whole cell lysate (20 µg)

    Lanes 1 - 3: Merged signal (red and green). Green - ab86808 observed at 24 kDa. Red - loading control, ab181602, observed at 37 kDa.

    Ab86808 was shown to specifically react with PGP9.5/UCHL1 in wild-type cells as signal was lost in PGP9/UCHL1 knockout HAP1 cells. Wild-type and UCHL1 (KO) knockout samples were subjected to SDS-PAGE.  Ab86808 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/2000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Overlay histogram showing SH-SH5Y cells stained with ab86808 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab86808, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in SH-SY5Y cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  • IHC-P image of PGP9.5 staining using ab86808(1:2500) on dog small intestine sections. The sections were subjected to heat mediated antigen retrieval using citric acid. The sections were then blocked with 1% BSA for 10 min at 21°C. the sections were then incubated with primary antibody ab86808 at 1:2500 dlution for 2 hours at 21°C. The secondary used was Goat polyclonal to anti mouse IgG conjugated to biotin (1:200)

    See Abreview

  • All lanes : Anti-PGP9.5 antibody [346CT2.5.1] (ab86808) at 1 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Brain (Rat) Tissue Lysate
    Lane 3 : Rat Cortex Tissue Lysate
    Lane 4 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate
    Lane 5 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
    Lane 6 : U-87 MG (Human glioblastoma astrocytoma) Whole Cell Lysate
    Lane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

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


    Exposure time : 1 minute
  • IHC-P image of PGP9.5 staining on rat pancreas using ab86808 (1:1000). The rat pancreatic sections were subjected to heat mediated antigen retrieval using citric acid. The sections were then blocked using 1% BSA at 21°C for 10 min. ab86808 was incubated at 21°C for 1 hours. The secondary antibody was Goat polyclonal to anti-mouse (rat pre-absorbed) conjugated to biotin (1:200).

    See Abreview

文献

ab86808 has not yet been referenced specifically in any publications.

客户评价及客户问答

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Goat Tissue sections (Small intestine)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Small intestine
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

提交于 Oct 14 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cat Tissue sections (Bladder)
Specification
Bladder
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

提交于 Nov 29 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (Small intestine)
Specification
Small intestine
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

提交于 Nov 15 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Pancreas)
Specification
Pancreas
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

提交于 Jun 13 2012

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

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