製品の概要

  • 製品名
    Anti-Proteasome 20S LMP7 antibody
    Proteasome 20S LMP7 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Proteasome 20S LMP7
  • 特異性
    Detects proteasome 20S LMP7.
  • アプリケーション
    適用あり: WB, IHC-P, ICC/IF, IHC-Fr, Flow Cytmore details
  • 種交差性
    交差種: Mouse, Rat, Human
    交差が予測される動物種: Sheep, Cow
  • 免疫原

    Synthetic peptide corresponding to Human Proteasome 20S LMP7 aa 259-274.
    Sequence:

    VESTDVSDLLHQYREA


    Database link: P28062
    (Peptide available as ab4945)

製品の特性

  • 製品の状態
    Liquid
  • 保存方法
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • バッファー
    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...
  • 精製度
    Immunogen affinity purified
  • ポリ/モノ
    ポリクローナル
  • アイソタイプ
    IgG
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab3329 in the following tested applications.

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

アプリケーション Abreviews 特記事項
WB Use a concentration of 1 - 3 µg/ml. Detects a band of approximately 20 kDa.Can be blocked with Human Proteasome 20S LMP7 peptide (ab4945).
IHC-P Use a concentration of 1 µg/ml.
ICC/IF Use a concentration of 5 µg/ml.
IHC-Fr 1/500. PubMed: 17008387
Flow Cyt Use 3-5µg for 106 cells.

ターゲット情報

  • 機能
    The proteasome is a multicatalytic proteinase complex which is characterized by its ability to cleave peptides with Arg, Phe, Tyr, Leu, and Glu adjacent to the leaving group at neutral or slightly basic pH. The proteasome has an ATP-dependent proteolytic activity. This subunit is involved in antigen processing to generate class I binding peptides. Replacement of PSMB5 by PSMB8 increases the capacity of the immunoproteasome to cleave model peptides after hydrophobic and basic residues. Acts as a major component of interferon gamma-induced sensitivity. Plays a key role in apoptosis via the degradation of the apoptotic inhibitor MCL1. May be involved in the inflammatory response pathway. In cancer cells, substitution of isoform 1 (E2) by isoform 2 (E1) results in immunoproteasome deficiency.
  • 関連疾患
    Defects in PSMB8 are the cause of JMP syndrome (JMPS) [MIM:613732]; also called joint contractures muscular atrophy microcytic anemia and panniculitis-induced lipodystrophy. JBTS1 is an autoinflammatory disorder characterized by childhood onset of joint stiffness and severe contractures of the hands and feet, erythematous skin lesions with subsequent development of severe lipodystrophy, and laboratory evidence of immune dysregulation. Accompanying features include muscle weakness and atrophy, hepatosplenomegaly, and microcytic anemia.
  • 配列類似性
    Belongs to the peptidase T1B family.
  • 発生段階
    Highly expressed in immature dendritic cells (at protein level).
  • 翻訳後修飾
    Autocleaved. The resulting N-terminal Thr residue of the mature subunit is responsible for the nucleophile proteolytic activity.
  • 細胞内局在
    Cytoplasm. Nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • ALDD antibody
    • D6S216 antibody
    • D6S216E antibody
    • Large multifunctional peptidase 7 antibody
    • Large multifunctional protease 7 antibody
    • LMP 7 antibody
    • LMP7 antibody
    • Low molecular mass protein 7 antibody
    • Low molecular weight protein 7 antibody
    • Macropain subunit C13 antibody
    • MGC1491 antibody
    • Multicatalytic endopeptidase complex subunit C13 antibody
    • NKJO antibody
    • OTTHUMP00000062981 antibody
    • Protease component C13 antibody
    • Proteasome (prosome macropain) subunit beta type 8 antibody
    • Proteasome (prosome, macropain) subunit, beta type, 8 (large multifunctional peptidase 7) antibody
    • Proteasome beta 8 subunit antibody
    • Proteasome catalytic subunit 3i antibody
    • Proteasome component C13 antibody
    • Proteasome related gene 7 antibody
    • Proteasome subunit beta 5i antibody
    • Proteasome subunit beta 8 antibody
    • Proteasome subunit beta type 8 antibody
    • Proteasome subunit beta type antibody
    • Proteasome subunit beta type-8 antibody
    • Proteasome subunit beta-5i antibody
    • Proteasome subunit Y2 antibody
    • PSB8_HUMAN antibody
    • PSMB 8 antibody
    • PSMB5i antibody
    • PSMB8 antibody
    • Really interesting new gene 10 protein antibody
    • RING 10 antibody
    • RING10 antibody
    • Y2 antibody
    see all

画像

  • All lanes : Anti-Proteasome 20S LMP7 antibody (ab3329) at 2 µg/ml

    Lane 1 : HeLa whole cell extracts
    Lane 2 : HeLa treated with IFN gamma (100ng/ml IFN gamma for 72h) whole cell extracts
    Lane 3 : U-937 whole cell extracts
    Lane 4 : Raji whole cell extracts
    Lane 5 : Ramos whole cell extracts

    Lysates/proteins at 30 µg per lane.

    Secondary
    Goat anti-Rabbit IgG (H+L) HRP conjugate at 1/2500 dilution

    Observed band size : 20 kDa (why is the actual band size different from the predicted?)

    Detected by chemiluminescence.

  • Immunocytochemistry/ Immunofluorescence analysis of HeLa cells labeling Proteasome 20S LMP7 with ab3329 at 5µg/ml. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 in TBS for 10 minutes, and blocked with 3% Blocker BSA in PBS for 15 minutes at room temperature. Cells were stained with or without Anti-Proteasome 20S LMP7 antibody (ab3329), at a concentration of 5µg/ml for 1 hour at room temperature, and then incubated with a Alexa Fluor® 488 goat anti-rabbit IgG secondary antibody at a dilution of 1/1000 for 1 hour s at room temperature (both panels, green). Nuclei (both panels, blue) were stained with Hoechst 33342 dye.

  • Ab3329 staining Human normal skin. Staining is localized to cytoplasmic and nuclear compartments.
    Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required
  • Flow Cytometry analysis of SH-SY5Y cells labeling Proteasome 20S LMP7 with ab3329. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Anti-Proteasome 20S LMP7 antibody (ab3329) (red histogram) or with rabbit isotype control (pink histogram) at 3-5 ug/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody at a dilution of 1/400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

  • Western blot of proteasome 20S LMP7 from HeLa cell extract using ab3329.
  • ICC/IF image of ab3329 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab3329, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

参考文献

This product has been referenced in:
  • Textor A  et al. Preventing tumor escape by targeting a post-proteasomal trimming independent epitope. J Exp Med 213:2333-2348 (2016). Read more (PubMed: 27697836) »
  • Rouette A  et al. Expression of immunoproteasome genes is regulated by cell-intrinsic and -extrinsic factors in human cancers. Sci Rep 6:34019 (2016). WB ; Human . Read more (PubMed: 27659694) »

See all 21 Publications for this product

レビューと Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Feb 29 2016

Application
Western blot
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Specification
Skeletal muscle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Feb 29 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Skeletal muscle)
Loading amount
30 µg
Specification
Skeletal muscle
Gel Running Conditions
Reduced Denaturing (15% gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Dr. G.K Sakellariou

Verified customer

投稿 May 15 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Recombinant protein (Recombinant LMP7-GST)
Loading amount
0.025 µg
Specification
Recombinant LMP7-GST
Gel Running Conditions
Reduced Denaturing (4-12% bis/tris)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

投稿 May 31 2012

Thank you for sending me this image. This data will be used to keep records of how our products are performing and for internal invetigation if needed. I noted that all tissue lysates that you have used have given the same non-specific signal. However ...

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Thank you for taking time to send us your protocols. I am sorry to hear that this antibody is not providing satisfactory results. The details provided will enable us to investigate this case and will provide us with vital information for monitoring pro...

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Thank you for contacting us. I would like to ask you if you might send me the protocols used with these productsso that we can gather further information regarding the samples tested and the protocol used. Once we receive your response, we will look at...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (dendritic cell)
Loading amount
1e+006 cells
Specification
dendritic cell
Gel Running Conditions
Non-reduced Denaturing
Blocking step
BSA as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Username

Abcam user community

Verified customer

投稿 Jan 05 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Purified protein (heart and spleen)
Loading amount
1 µg
Specification
heart and spleen
Gel Running Conditions
Reduced Denaturing (12,5%)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

投稿 Sep 03 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Cell lysate - whole cell (D1 cells)
Loading amount
35 µg
Specification
D1 cells
Gel Running Conditions
Reduced Denaturing
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

投稿 Mar 06 2008

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

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