Overview

  • Product name

    Anti-Cyclophilin B antibody
    See all Cyclophilin B primary antibodies
  • Description

    Rabbit polyclonal to Cyclophilin B
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IHC-Fr, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Horse, Chicken, Dog, Human
    Predicted to work with: Cow, Pig, Xenopus laevis
  • Immunogen

    Synthetic peptide corresponding to Human Cyclophilin B aa 150 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab16277, ab5016)

  • Positive control

    • WB: HeLa, HeLa Nuclear, Jurkat, A431, HEK293, NIH 3T3, HAP1 and U87-MG cell lysates; Rat Liver Tissue IF: NIH3T3 cells

Properties

Applications

Our Abpromise guarantee covers the use of ab16045 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 Use a concentration of 0.5 µg/ml. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).Can be blocked with Human Cyclophilin B peptide (ab16277).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
ICC/IF Use a concentration of 1 µg/ml.
IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.

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 (ab16045) at 1/5000 dilution

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



    Lanes 1 - 4: Merged signal (red and green). Green - ab16045 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab16045 was shown to specifically react with PPIB in wild-type HAP1 cells as signal was lost in PPIB knockout cells. Wild-type and PPIB knockout samples were subjected to SDS-PAGE. Ab16045 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution 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.

  • ab16045 stained HeLa cells. The cells were 100% methanol fixed for 5 minutes at -20°C and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1hour at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab16045 at 5µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • All lanes : Anti-Cyclophilin B antibody (ab16045) at 1 µg/ml

    Lane 1 : Rat Liver
    Lane 2 : Mouse 3T3
    Lane 3 : Dog
    Lane 4 : Chicken

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 21 kDa
    Observed band size: 25 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds
  • All lanes : Anti-Cyclophilin B antibody (ab16045) at 1 µg/ml

    Lane 1 : HeLa nuclear lysate
    Lane 2 : HeLa whole cell lysate
    Lane 3 : A431 whole cell lysate
    Lane 4 : Jurkat whole cell lysate
    Lane 5 : HEK293 whole cell lysate
    Lane 6 : HeLa nuclear lysate with Human Cyclophilin B peptide (ab16277) at 1 µg/ml
    Lane 7 : HeLa whole cell lysate with Human Cyclophilin B peptide (ab16277) at 1 µg/ml
    Lane 8 : A431 whole cell lysate with Human Cyclophilin B peptide (ab16277) at 1 µg/ml
    Lane 9 : Jurkat whole cell lysate with Human Cyclophilin B peptide (ab16277) at 1 µg
    Lane 10 : HEK293 whole cell lysate with Human Cyclophilin B peptide (ab16277) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size: 21 kDa
    Observed band size: 21 kDa


    Exposure time: 30 seconds
  • Cyclophilin B was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to Cyclophilin B and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab16045.
    Secondary: Clean blot (HRP conjugate) at 1/1000 dilution.
    Band: 21kDa: Cyclophilin B.
  • ICC/IF image of ab16045 stained NIH/3T3 cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab16045, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • ab16045 (1/1000) staining Cyclophilin B in assynchronous HeLa cells (green). Cells were fixed with Paraformaldehyde and counter-stained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.

    See Abreview

  • Image courtesy of Human Protein Atlas

    ab16045 staining in human placenta, showing staining of the cytrotrophoblasts. Paraffin embedded placental tissue was incubated with ab16045 (1:14,000 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab16045 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org

    .

  • All lanes : Anti-Cyclophilin B antibody (ab16045) at 1/1000 dilution

    Lane 1 : Whole cell lysate prepared from rat pancreatic AR42J cells, which were treated with 10nM dexamethasone for 48 hours.
    Lane 2 : Whole cell lysate for negative control, prepared from rat pancreatic AR42J cells (specific knock down of cyclophilin B/PpiB by siRNA), which were treated with 10nM dexamethasone for 48 hours.

    Secondary
    All lanes : Goat-anti-Rabbit HRP-conjugated polyclonal at 1/2000 dilution

    Developed using the ECL technique.

    Predicted band size: 21 kDa
    Observed band size: 23 kDa why is the actual band size different from the predicted?


    Exposure time: 1 minute


    Primary antibody incubated for 12 hours at 4°C.
    Blocking step performed using 5% milk, 1 hour at 20°C.

    See Abreview

  • Anti-Cyclophilin B antibody (ab16045) at 0.5 µg/ml + Recombinant Human Cyclophilin B protein (ab88801) at 0.01 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 21 kDa


    Exposure time: 30 seconds

References

This product has been referenced in:

  • Chavez RD  et al. Prg4 prevents osteoarthritis induced by dominant-negative interference of TGF-ß signaling in mice. PLoS One 14:e0210601 (2019). Read more (PubMed: 30629676) »
  • Desmarais F  et al. Apolipoprotein D overexpression alters hepatic prostaglandin and omega fatty acid metabolism during the development of a non-inflammatory hepatic steatosis. Biochim Biophys Acta Mol Cell Biol Lipids 1864:522-531 (2019). Read more (PubMed: 30630053) »
See all 57 Publications for this product

Customer reviews and Q&As

Filter by Application

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1-8 of 8 Abreviews

Application
Western blot
Sample
Sheep Tissue lysate - whole (Placenta, caruncle)
Gel Running Conditions
Reduced Denaturing (10% polyacrylamide)
Loading amount
10 µg
Specification
Placenta, caruncle
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Miss. Barbara Makela

Verified customer

Submitted Oct 30 2015

Application
Immunocytochemistry/ Immunofluorescence
Sample
Horse Cell (fibroblasts)
Specification
fibroblasts
Fixative
Methanol
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C

Dr. Shawna Pyott

Verified customer

Submitted Aug 24 2012

Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Kidney)
Specification
Kidney
Fixative
Acetone
Permeabilization
No
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 21°C

Abcam user community

Verified customer

Submitted Mar 02 2012

Application
Western blot
Sample
Rat Cell lysate - whole cell (pancreatic AR42J cells)
Loading amount
20 µg
Specification
pancreatic AR42J cells
Treatment
10 nM dexamethasone for 48 hrs
Gel Running Conditions
Reduced Denaturing (12.5%, 5% stacking gel)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Dr. Michael Schrader

Verified customer

Submitted Sep 16 2010

Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (rat exocrine pancreas)
Specification
rat exocrine pancreas
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 4% · Temperature: 20°C

Dr. Michael Schrader

Verified customer

Submitted Sep 15 2010

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (panceatic AR42J cells)
Specification
panceatic AR42J cells
Fixative
Paraformaldehyde
Permeabilization
Yes - Triton X-100
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 20°C

Dr. Michael Schrader

Verified customer

Submitted Sep 14 2010

Application
Western blot
Sample
Rat Tissue lysate - whole (Kidney)
Loading amount
20 µg
Specification
Kidney
Gel Running Conditions
Reduced Denaturing (10% Bis Tris Gels)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Mar 19 2010

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Hela)
Specification
Hela
Fixative
Paraformaldehyde
Permeabilization
Yes - PBS TritonX100 (0.5%;10min)

Dr. Kirk Mcmanus

Verified customer

Submitted Jun 28 2007

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