Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)

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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
  • Suitable for: Flow Cyt, IHC-P, WB, ICC/IF
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Proteasome 20S LMP7 antibody [EPR14482(B)]
    See all Proteasome 20S LMP7 primary antibodies

  • Description

    Rabbit monoclonal [EPR14482(B)] to Proteasome 20S LMP7

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, IHC-P, WB, ICC/IFmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide within Human Proteasome 20S LMP7 aa 200 to the C-terminus. The exact sequence is proprietary.
    Database link: P28062

  • Positive control

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

  • General notes

    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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR14482(B)

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab180606 in the following tested applications.

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

Application Abreviews Notes
Flow Cyt 1/90 - 1/150.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 
IHC-P 1/1700.

For unpurified use at 1/250 - 1/500.

Perform heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0)
WB 1/10000 - 1/50000. Detects a band of approximately 23 kDa (predicted molecular weight: 30 kDa).
ICC/IF 1/100.
  • Application notes
    Is unsuitable for IP.

    • Target

    • Function

      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.

    • Involvement in disease

      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.

    • Sequence similarities

      Belongs to the peptidase T1B family.

    • Developmental stage

      Highly expressed in immature dendritic cells (at protein level).

    • Post-translational
      modifications

      Autocleaved. The resulting N-terminal Thr residue of the mature subunit is responsible for the nucleophile proteolytic activity.

    • Cellular localization

      Cytoplasm. Nucleus.
    • Information by UniProt

    • Database links

      • Alternative names

        • 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

      Images

      • Western blot - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)
        Western blot - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)
        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?



        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)] (ab180606)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)

        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.

      • Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)
        Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)
        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.
      • Flow Cytometry - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)
        Flow Cytometry - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)
        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).
      • Western blot - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)
        Western blot - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)
        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?



        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.

      • Western blot - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)
        Western blot - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)
        Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606) at 1/10000 dilution (Purified) + Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysates at 15 µg

        Secondary
        Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)

        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.

      • Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)
        Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)

        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.

      • Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)
        Immunocytochemistry/ Immunofluorescence - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)

        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).

      • Flow Cytometry - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)
        Flow Cytometry - Anti-Proteasome 20S LMP7 antibody [EPR14482(B)] (ab180606)

        Flow cytometric analysis of 2% paraformaldehyde-fixed Jurkat (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 by Goat anti rabbit IgG (FITC) secondary antibody at 1/150 dilution.

      References (2)

      Publishing research using ab180606? Please let us know so that we can cite the reference in this datasheet.

      ab180606 has been referenced in 2 publications.

      • Javitt A  et al. Pro-inflammatory Cytokines Alter the Immunopeptidome Landscape by Modulation of HLA-B Expression. Front Immunol 10:141 (2019). PubMed: 30833945
      • Xie X  et al. The immunoproteasome catalytic ß5i subunit regulates cardiac hypertrophy by targeting the autophagy protein ATG5 for degradation. Sci Adv 5:eaau0495 (2019). PubMed: 31086810

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