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遺伝子ノックアウト細胞株 検証済

Anti-PGP9.5 抗体 [31A3] (ab20559)

Submit a review Q&A (1)References (8)
Western blot - Anti-PGP9.5 antibody [31A3] (ab20559)
  • Flow Cytometry - Anti-PGP9.5 antibody [31A3] (ab20559)
  • Western blot - Anti-PGP9.5 antibody [31A3] (ab20559)

Key features and details

  • Mouse monoclonal [31A3] to PGP9.5
  • Suitable for: Flow Cyt, WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-PGP9.5 antibody [31A3]
    PGP9.5 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [31A3] to PGP9.5

  • 由来種

    Mouse

  • アプリケーション

    適用あり: Flow Cyt, WBmore details

  • 種交差性

    交差種: Mouse, Rat, Human

  • 免疫原

    Full length protein corresponding to PGP9.5.

  • 特記事項

    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.

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab20559の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
Flow Cyt
Use 0.1-1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 
WB
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).

Dilution optimised using Chromogenic detection.
特記事項
Flow Cyt
Use 0.1-1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 
WB
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 25 kDa (predicted molecular weight: 25 kDa).

Dilution optimised using Chromogenic detection.

ターゲット情報

  • 機能

    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.

  • 組織特異性

    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.

  • 関連疾患

    Parkinson disease 5
    Neurodegeneration with optic atrophy, childhood-onset

  • 配列類似性

    Belongs to the peptidase C12 family.

  • 翻訳後修飾

    O-glycosylated.

  • 細胞内局在

    Cytoplasm. Endoplasmic reticulum membrane. About 30% of total UCHL1 is associated with membranes in brain.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • 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

画像

  • Western blot - Anti-PGP9.5 antibody [31A3] (ab20559)
    Western blot - Anti-PGP9.5 antibody [31A3] (ab20559)

    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)
    Lane 4: Hek293 whole cell lysate (20 µg)              

    Lanes 1 - 4: Merged signal (red and green). Green - ab20559 observed at 24 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab20559 detected the expected band for PGP9.5 in wild type cells and the band was not seen in PGP9.5/UCHL1 knockout cells. Wild-type and UCHL1 knockout samples were subjected to SDS-PAGE.  Ab20559 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 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.

  • Flow Cytometry - Anti-PGP9.5 antibody [31A3] (ab20559)
    Flow Cytometry - Anti-PGP9.5 antibody [31A3] (ab20559)
    Overlay histogram showing SH-SH5Y cells stained with ab20559 (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 (ab20559, 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.
  • Western blot - Anti-PGP9.5 antibody [31A3] (ab20559)
    Western blot - Anti-PGP9.5 antibody [31A3] (ab20559)
    All lanes : Anti-PGP9.5 antibody [31A3] (ab20559) at 5 µg/ml

    Lane 1 : Human brain tissue lysate - total protein (ab29466)
    Lane 2 : Human spinal cord tissue lysate - total protein (ab29188)
    Lane 3 : Brain (Rat) Tissue Lysate
    Lane 4 : Brain (Mouse) Tissue Lysate
    Lane 5 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 25 kDa
    Additional bands at: 25 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 4 minutes

参考文献 (8)

ab20559 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab20559 は 8 報の論文で使用されています。

  • Mtango NR  et al. Essential role of maternal UCHL1 and UCHL3 in fertilization and preimplantation embryo development. J Cell Physiol 227:1592-603 (2012). ICC/IF ; Mouse . PubMed: 21678411
  • Saygili E  et al. Age-related regional differences in cardiac nerve growth factor expression. Age (Dordr) 34:659-67 (2012). WB ; Rat . PubMed: 21559866
  • Mtango NR  et al. Essential role of ubiquitin C-terminal hydrolases UCHL1 and UCHL3 in mammalian oocyte maturation. J Cell Physiol 227:2022-9 (2012). PubMed: 21751213
  • Wang W  et al. Abnormal innervation patterns in the anorectum of ETU-induced fetal rats with anorectal malformations. Neurosci Lett 495:88-92 (2011). IHC-P ; Rat . PubMed: 21440597
  • Zippin JH  et al. Soluble adenylyl cyclase defines a nuclear cAMP microdomain in keratinocyte hyperproliferative skin diseases. J Invest Dermatol 130:1279-87 (2010). IHC-P ; Human . PubMed: 20130594
  • Rana OR  et al. Acetylcholine as an age-dependent non-neuronal source in the heart. Auton Neurosci 156:82-9 (2010). WB ; Rat . PubMed: 20510655
  • Crimmins S  et al. Transgenic rescue of ataxia mice reveals a male-specific sterility defect. Dev Biol 325:33-42 (2009). IHC-P ; Mouse . PubMed: 18926813
  • Yi YJ  et al. Ubiquitin C-terminal hydrolase-activity is involved in sperm acrosomal function and anti-polyspermy defense during porcine fertilization. Biol Reprod 77:780-93 (2007). IHC-P ; Mouse . PubMed: 17671268

レビューと Q&A

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レビューを送る 質問を送る

Question

Hi, I would like to know if you could recommend a ELISA antibody pair for PGP9.5 (UCH-L1). I am looking for a good pair of antibodies for kinetic sandwich ELISA assay for UCH-L1. Due to the nature of the assay, I need antibodies with high affinity and fast on-rate. Could you please recommend your highest affinity antibodies for UCH-L1? Thanks,  

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Answer

Thank you for contacting us. We have two anti-PGP9.5 antibodies in our catalogue which have been tested using ELISA which I can suggest to you. ab20559 is a mouse monoclonal, reactive against human and rat PGP9.5 and ab53057 is a rabbit polyclonal reactive against human  PGP9.5 (and is predicted to react with rat protein). Whilst these have been used in ELISAs before, they have not to our knowledge been used as a pair for a sandwich ELISA. Unfortunately we do not have any information for either antibody about the affinity or the "on-rate". I am sorry that I could not be of more help. If you require any further information please do not hesitate to ask.

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