Validated using a knockout cell line
Recombinant
RabMAb

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

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

    Suitable for: WBmore details
    Unsuitable for: Flow Cyt,ICC/IF or IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • 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

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab178397 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 1/1000 - 1/10000. Predicted molecular weight: 24 kDa.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IHC-P.
  • Target

    • Function

      PPIases accelerate the folding of proteins. It catalyzes the cis-trans isomerization of proline imidic peptide bonds in oligopeptides.
    • Involvement in disease

      Defects in PPIB are the cause of osteogenesis imperfecta type 9 (OI9) [MIM:259440]. OI9 is a connective tissue disorder characterized by bone fragility, low bone mass and bowing of limbs due to multiple fractures. Short limb dwarfism and blue sclerae are observed in some but not all patients.
    • Sequence similarities

      Belongs to the cyclophilin-type PPIase family. PPIase B subfamily.
      Contains 1 PPIase cyclophilin-type domain.
    • Cellular localization

      Endoplasmic reticulum lumen. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
    • Information by UniProt
    • Database links

    • Alternative names

      • AA408962 antibody
      • AA553318 antibody
      • AI844835 antibody
      • Cphn 2 antibody
      • Cphn2 antibody
      • Cyclophilin B antibody
      • Cyclophilin like protein antibody
      • CyP 20b antibody
      • CYP S1 antibody
      • CYP-S1 antibody
      • CYPB antibody
      • EC 5.2.1.8 antibody
      • MGC14109 antibody
      • MGC2224 antibody
      • OI9 antibody
      • peptidyl prolyl cis trans isomerase B antibody
      • Peptidyl prolyl cis trans isomerase B precursor antibody
      • Peptidyl-prolyl cis-trans isomerase B antibody
      • peptidylprolyl isomerase B (cyclophilin B) antibody
      • Peptidylprolyl isomerase B antibody
      • PPIase antibody
      • PPIase B antibody
      • Ppib antibody
      • PPIB_HUMAN antibody
      • Rotamase antibody
      • Rotamase B antibody
      • S cyclophilin antibody
      • S-cyclophilin antibody
      • SCYLP antibody
      see all

    Images

    • 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.

    • 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.

    • 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.

    • 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

    References

    This product has been referenced in:

    • Lee E  et al. GREB1 amplifies androgen receptor output in human prostate cancer and contributes to antiandrogen resistance. Elife 8:N/A (2019). Read more (PubMed: 30644358) »
    • Mann V  et al. Changes in Human Foetal Osteoblasts Exposed to the Random Positioning Machine and Bone Construct Tissue Engineering. Int J Mol Sci 20:N/A (2019). Read more (PubMed: 30889841) »
    See all 6 Publications for this product

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