Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

Overview

  • Product name

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

  • Description

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

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • 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

    Ab246363 is the carrier-free version of ab180606. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab246363 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    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.

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.

  • Storage buffer

    pH: 7.2
    Constituent: PBS

  • Carrier free

    Yes
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR14482(B)

  • Isotype

    IgG

  • Research areas

Images

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

    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.

    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)

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