Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR12703(B)] to Cyclophilin B - Loading Control
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control
    See all Cyclophilin B primary antibodies

  • Description

    Rabbit monoclonal [EPR12703(B)] to Cyclophilin B - Loading Control

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    WB
    Mouse
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human Cyclophilin B aa 150 to the C-terminus (Cysteine residue). The exact sequence is proprietary.
    Database link: P23284

  • Positive control

    • WB: HepG2, HeLa, NIH/3T3, PC-12, HAP1, Jurkat, U87-MG and A431 cell lysates, mouse and rat heart tissue lysates.

  • 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.

Properties

Images

  • Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)
    Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)
    All lanes : Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397) at 1/1000 dilution

    Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : PPIB knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate
    Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) 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: 24 kDa
    Observed band size: 24 kDa



    Lanes 1-4: Merged signal (red and green). Green - ab178397 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab178397 Anti-Cyclophilin B antibody [EPR12703(B)] was shown to specifically react with Cyclophilin B in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261746 (knockout cell lysate ab257037) was used. Wild-type and Cyclophilin B knockout samples were subjected to SDS-PAGE. ab178397 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-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)
    Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)
    All lanes : Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397) at 1 µg/ml

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : PPIB knockout HAP1 whole cell lysate
    Lane 3 : Jurkat whole cell lysate
    Lane 4 : U87-MG whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 24 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab178397 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab178397 was shown to recognize PPIB in wild-type HAP1 cells as signal was lost at the expected MW in PPIB knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and PPIB knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab178397 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)
    Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)
    All lanes : Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397) at 1/10000 dilution (purified)

    Lane 1 : Mouse heart tissue lysate
    Lane 2 : NIH/3T3 cell lysate
    Lane 3 : Rat heart tissue lysate
    Lane 4 : PC-12 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size: 24 kDa
    Observed band size: 18 kDa
    why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)
    Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)
    All lanes : Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397) at 1/10000 dilution (purified)

    Lane 1 : HepG2 cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : A431 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

    Predicted band size: 24 kDa
    Observed band size: 18 kDa why is the actual band size different from the predicted?



    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)
    Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)
    All lanes : Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397) at 1/1000 dilution

    Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
    Lane 3 : A431 (Human epidermoid carcinoma cell line) cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Developed using the ECL technique.

    Predicted band size: 24 kDa

  • Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)
    Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)

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