Overview

  • Product nameAnti-Cyclophilin B antibody [k2E2]
    See all Cyclophilin B primary antibodies
  • Description
    Mouse monoclonal [k2E2] to Cyclophilin B
  • SpecificityThis antibody does not cross react with Cyclophilin A, D, E, F, G and H.
  • Tested applicationsSuitable for: IHC-P, Flow Cyt, WB, ELISA, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant human Cyclophilin B (26-216 aa) purified from E. coli

  • Positive control
    • HepG2 and HeLa cell lysates.
  • General notes


    ab74173, clone k2E2, is derived from hybridization of mouse FO myeloma cells with spleen cells from BALB/c mice immunized with a recombinant human Cyclophilin B protein.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • Storage bufferPreservative: 0.1% Sodium Azide
    Constituents: PBS, pH 7.4
  • Concentration information loading...
  • PurityProtein G purified
  • Purification notesab74173 was purified from mouse ascitic fluids by protein G affinity chromatography.
  • Primary antibody notesab74173, clone k2E2, is derived from hybridization of mouse FO myeloma cells with spleen cells from BALB/c mice immunized with a recombinant human Cyclophilin B protein.
  • ClonalityMonoclonal
  • Clone numberk2E2
  • IsotypeIgG1
  • Light chain typekappa
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab74173 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
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
WB 1/1000 - 1/2000. Predicted molecular weight: 23 kDa.
ELISA Use at an assay dependent concentration.
ICC/IF Use a concentration of 1 µg/ml.

Target

  • FunctionPPIases accelerate the folding of proteins. It catalyzes the cis-trans isomerization of proline imidic peptide bonds in oligopeptides.
  • Involvement in diseaseDefects 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 similaritiesBelongs to the cyclophilin-type PPIase family. PPIase B subfamily.
    Contains 1 PPIase cyclophilin-type domain.
  • Cellular localizationEndoplasmic 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

Anti-Cyclophilin B antibody [k2E2] images

  • All lanes : Anti-Cyclophilin B antibody [k2E2] (ab74173) at 1/1000 dilution

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

    Lysates/proteins at 30 µg per lane.

    Secondary
    goat anti-mouse antibody conjugated to HRP
    Developed using the ECL technique

    Predicted band size : 23 kDa
    Observed band size : 21 kDa (why is the actual band size different from the predicted?)
  • ICC/IF image of ab74173 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab74173, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • IHC image of ab74173 staining in human normal liver formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH9, epitope retrieval solution 2) for 20 mins. The section was then incubated with ab74173, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • Overlay histogram showing HeLa cells stained with ab74173 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab74173, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Hela cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References for Anti-Cyclophilin B antibody [k2E2] (ab74173)

This product has been referenced in:

See all 2 Publications for this product

Product Wall

Application Western blot
Sample Rat Tissue lysate - whole (Whole Olfactory epitherlium)
Gel Running Conditions Reduced Denaturing (4-20% Tris Gly Gel)
Loading amount 25 µg
Specification Whole Olfactory epitherlium
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Ms. Misha Riley

Verified customer

Submitted Jun 22 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (kidney)
Loading amount 20 µg
Specification kidney
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Nov 30 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"