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遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

Anti-Proteasome 20S LMP7 抗体 [EPR14482(B)] - BSA and Azide free (ab246363)

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Western blot - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
  • Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
  • Flow Cytometry (Intracellular) - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
  • Western blot - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
  • Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
  • Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
  • Flow Cytometry (Intracellular) - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
  • Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR14482(B)] to Proteasome 20S LMP7 - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 PE Unconjugated

製品の概要

  • 製品名

    Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free
    Proteasome 20S LMP7 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR14482(B)] to Proteasome 20S LMP7 - BSA and Azide free

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
    適用なし: IP

  • 種交差性

    交差種: Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: A549, Raji, Jurkat, HeLa and U937 lysates. IHC-P: Human bladder transitional cell carcinoma tissue. ICC/IF: Jurkat and HeLa cells. Flow Cyt (intra): Jurkat cells. IP: Proteasome 20S LMP7 IP in Raji cell lysate.

  • 特記事項

    ab246363 is the carrier-free version of ab180606.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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ab246363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 (Intra)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 23 kDa (predicted molecular weight: 30 kDa).
IHC-P
Use at an assay dependent concentration.

Perform heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0).
ICC/IF
Use at an assay dependent concentration.
特記事項
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 23 kDa (predicted molecular weight: 30 kDa).
IHC-P
Use at an assay dependent concentration.

Perform heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0).
ICC/IF
Use at an assay dependent concentration.
追加情報
Is unsuitable for IP.

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

    • Western blot - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
      Western blot - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
      All lanes : Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606) at 1/1000 dilution

      Lane 1 : Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
      Lane 2 : PSMB8 knockout A549 (Human lung carcinoma cell line) whole cell lysate
      Lane 3 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
      Lane 4 : HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 30 kDa
      Observed band size: 23 kDa why is the actual band size different from the predicted?



      This data was developed using ab180606, the same antibody clone in a different buffer formulation.

      Lanes 1-4: Merged signal (red and green). Green - ab180606 observed at 23 kDa. Red - loading control ab8245 observed at 36 kDa.

       ab180606 Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] was shown to specifically react with Proteasome 20S LMP7 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267149 (knockout cell lysate ab257130) was used. Wild-type and Proteasome 20S LMP7 knockout samples were subjected to SDS-PAGE. ab180606 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric carcinoma tissue sections labeling Proteasome 20S LMP7 with purified ab180606 at 1/1700 dilution (0.52 µg/ml). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab180606)

    • Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
      Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
      Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling Proteasome 20S LMP7 with purified ab180606 at 1:100 dilution (8.9 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (anti proteasome 20s lmp7 antibody epr14482 b immunocytochemistry hela human)
    • Flow Cytometry (Intracellular) - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
      Flow Cytometry (Intracellular) - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)

      Intracellular Flow Cytometry analysis of Raji (Human Burkitt's lymphoma B lymphocyte) cells labeling Proteasome 20S LMP7 with purified ab180606 at 1/90 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab180606)

       

    • Western blot - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
      Western blot - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
      All lanes : Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606) at 1/1000 dilution

      Lane 1 : Wild-type A549 cell lysate
      Lane 2 : PSMB8 knockout A549 cell lysate
      Lane 3 : Raji cell lysate
      Lane 4 : HEK-293 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

      Predicted band size: 30 kDa
      Observed band size: 23 kDa why is the actual band size different from the predicted?



      This data was developed using the same antibody clone in a different buffer formulation (ab180606).

      Lanes 1-4: Merged signal (red and green). Green - ab180606 observed at 23 kDa. Red - loading control ab8245 observed at 36 kDa.

       ab180606 Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] was shown to specifically react with Proteasome 20S LMP7 in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267148 (knockout cell lysate ab257129) was used. Wild-type and Proteasome 20S LMP7 knockout samples were subjected to SDS-PAGE. ab180606 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)

      Immunohistochemical analysis of paraffin-embedded human bladder transitional cell carcinoma tissue labeling Proteasome 20S LMP7 with ab180606 (unpurified at 1/500 dilution, followed by prediluted ImmunoHistoprobe (Ready to use) HRP Polymer for Rabbit IgG. Counterstained with hematoxylin.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab180606).

    • Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
      Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)

      ab180606 (unpurified) staining Proteasome 20S LMP7 in Jurkat (human acute T cell leukemia) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used as a nuclear counterstain.

      Negative control: PBS only.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab180606).

    • Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
      Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)

      Immunofluorecenct analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Proteasome 20S LMP7 with ab180606 (unpurified at 1/100 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody at 1/200 dilution (left panel). DAPI staining (right panel).

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab180606).

    • Flow Cytometry (Intracellular) - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
      Flow Cytometry (Intracellular) - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)

      Intracellular flow cytometric analysis of 2% paraformaldehyde-fixedJurkat (Human T cell leukemia cell line from peripheral blood) cells labeling Proteasome 20S LMP7 with ab180606 (unpurified at 1/150 dilution (red) compared to a Rabbit monoclonal IgG Isotype control (green), followed byGoat anti rabbit IgG (FITC) secondary antibody at 1/150 dilution. 

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol and sodium azide (ab180606). 

       

       

    • Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)
      Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] - BSA and Azide free (ab246363)

    プロトコール

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    参考文献 (0)

    ab246363 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。

    ab246363 は論文での使用が確認できていません。

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