Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-CSN7b antibody [EPR6464] (ab133548)

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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 [EPR6464] to CSN7b
  • Suitable for: WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-CSN7b antibody [EPR6464]
    See all CSN7b primary antibodies

  • Description

    Rabbit monoclonal [EPR6464] to CSN7b

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, Flow Cytmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human CSN7b aa 50-150. The exact sequence is proprietary.

  • Positive control

    • WB: HEK293T, HAP1, Jurkat and HeLa cell lysates. IHC-P: Human kidney tissue.

  • 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab133548 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
WB 1/10000 - 1/50000. Detects a band of approximately 30 kDa (predicted molecular weight: 30 kDa).
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Flow Cyt 1/200.
  • Application notes
    Is unsuitable for IP.

    • Target

    • Function

      Component of the COP9 signalosome complex (CSN), a complex involved in various cellular and developmental processes. The CSN complex is an essential regulator of the ubiquitin (Ubl) conjugation pathway by mediating the deneddylation of the cullin subunits of SCF-type E3 ligase complexes, leading to decrease the Ubl ligase activity of SCF-type complexes such as SCF, CSA or DDB2. The complex is also involved in phosphorylation of p53/TP53, JUN, I-kappa-B-alpha/NFKBIA, ITPK1 and IRF8/ICSBP, possibly via its association with CK2 and PKD kinases. CSN-dependent phosphorylation of TP53 and JUN promotes and protects degradation by the Ubl system, respectively.

    • Sequence similarities

      Belongs to the CSN7/EIF3M family. CSN7 subfamily.
      Contains 1 PCI domain.

    • Cellular localization

      Cytoplasm. Nucleus.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • COP9 Complex Homolog Subunit 7b antibody
      • COP9 Constitutive Photomorphogenic Homolog Subunit 7b antibody
      • COP9 Signalasome Subunit 7b antibody
      • COP9 signalosome complex subunit 7b antibody
      • COPS7B antibody
      • CSN7B_HUMAN antibody
      • JAB1 Containing Signalosome Subunit 7b antibody
      • JAB1-containing signalosome subunit 7b antibody
      • SGN7b antibody
      • Signalosome subunit 7b antibody
      see all

    Images

    • Western blot - Anti-CSN7b antibody [EPR6464] (ab133548)
      Western blot - Anti-CSN7b antibody [EPR6464] (ab133548)
      All lanes : Anti-CSN7b antibody [EPR6464] (ab133548) at 1/1000 dilution

      Lane 1 : Wild-type HEK293T cell lysate
      Lane 2 : COPS7B knockout HEK293T cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : HAP1 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: 32 kDa
      why is the actual band size different from the predicted?



      Lanes 1-4: Merged signal (red and green). Green - ab133548 observed at 32 kDa. Red - loading control ab8245 observed at 36 kDa. 

       ab133548 Anti-CSN7b antibody [EPR6464] was shown to specifically react with CSN7b in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266646 (knockout cell lysate ab257895) was used.  Wild-type and CSN7b knockout samples were subjected to SDS-PAGE.  ab133548 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-CSN7b antibody [EPR6464] (ab133548)
      Western blot - Anti-CSN7b antibody [EPR6464] (ab133548)

      Lane 1: Wild-type HAP1 cell lysate (20 µg)
      Lane 2: CSN7b knockout HAP1 cell lysate (20 µg)
      Lane 3: HeLa cell lysate (20 µg)
      Lanes 1 - 3: Merged signal (red and green). Green - ab133548 observed at 32 kDa. Red - loading control, ab18058, observed at 124 kDa.
      ab133548 was shown to specifically react with CSN7b when CSN7b knockout samples were used. Wild-type and CSN7b knockout samples were subjected to SDS-PAGE. ab133548 and ab18058 (loading control to vinculin) were both diluted 1/10000 and incubated overnight at 4°C. 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 1/10000 dilution for 1 hour at room temperature before imaging.

    • Flow Cytometry - Anti-CSN7b antibody [EPR6464] (ab133548)
      Flow Cytometry - Anti-CSN7b antibody [EPR6464] (ab133548)

      Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling CSN7b with unpurified ab133548 at 1/200 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti-rabbit IgG (Alexa Fluor® 488)(1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CSN7b antibody [EPR6464] (ab133548)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CSN7b antibody [EPR6464] (ab133548)

      Immunohistochemical analysis of paraffin embedded Human kidney tissue labelling CSN7b with ab133548 antibody at a dilution of 1/250.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Western blot - Anti-CSN7b antibody [EPR6464] (ab133548)
      Western blot - Anti-CSN7b antibody [EPR6464] (ab133548)
      All lanes : Anti-CSN7b antibody [EPR6464] (ab133548) at 1/10000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : HeLa cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 30 kDa
      Observed band size: 30 kDa

    • Other - Anti-CSN7b antibody [EPR6464] (ab133548)
      Other - Anti-CSN7b antibody [EPR6464] (ab133548)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    Publishing research using ab133548? Please let us know so that we can cite the reference in this datasheet.

    ab133548 has been referenced in 1 publication.

    • Liu J  et al. A genome-scale CRISPR-Cas9 screening in myeloma cells identifies regulators of immunomodulatory drug sensitivity. Leukemia N/A:N/A (2018). PubMed: 30026574

    Customer reviews and Q&As

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    Western blot abreview for Anti-CSN7b antibody [EPR6464]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HAP1 cell line)
    Gel Running Conditions
    Non-reduced Denaturing (4-12% Bis-Tris gel Bis-Tris gel)
    Loading amount
    50 µg
    Specification
    HAP1 cell line
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More

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    Verified customer

    Submitted Oct 10 2016

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