- Datasheet
- Specific References (39)
- Protocols
Overview
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Product nameAnti-Cyclophilin B antibody
See all Cyclophilin B primary antibodies -
DescriptionRabbit polyclonal to Cyclophilin B
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Host speciesRabbit
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Tested applicationsSuitable for: WB, IHC-P, ICC/IF, IHC-Fr, IPmore details
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Species reactivityReacts with: Mouse, Rat, Horse, Chicken, Dog, Human
Predicted to work with: Cow, Pig, Xenopus laevis
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human Cyclophilin B.
(Peptide available as ab16277.)
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Positive control
- This antibody gave a positive signal in: (WB) HeLa; HeLa Nuclear; Jurkat; A431; HEK293; NIH 3T3; Rat Liver Tissue: (IF) NIH3T3.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Immunohistochemistry kits
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Positive Controls
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
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FunctionPPIases accelerate the folding of proteins. It catalyzes the cis-trans isomerization of proline imidic peptide bonds in oligopeptides.
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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.
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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
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Database links
- Entrez Gene: 5479 Human
- Entrez Gene: 19035 Mouse
- Entrez Gene: 64367 Rat
- Omim: 123841 Human
- SwissProt: P23284 Human
- SwissProt: P24369 Mouse
- SwissProt: P24368 Rat
- Unigene: 434937 Human
see all -
Alternative names
- AA408962 antibody
- AA553318 antibody
- AI844835 antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin B antibody (ab16045)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.
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Western blot - Anti-Cyclophilin B antibody (ab16045)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 -
Western blot - Anti-Cyclophilin B antibody (ab16045)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 withHuman Cyclophilin B peptide (ab16277) at 1 µg/ml
Lane 7 : HeLa whole cell lysate withHuman Cyclophilin B peptide (ab16277) at 1 µg/ml
Lane 8 : A431 whole cell lysate withHuman Cyclophilin B peptide (ab16277) at 1 µg/ml
Lane 9 : Jurkat whole cell lysate withHuman Cyclophilin B peptide (ab16277) at 1 µg
Lane 10 : HEK293 whole cell lysate withHuman 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 -
Immunoprecipitation - Anti-Cyclophilin B antibody (ab16045)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. -
Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin B antibody (ab16045)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).
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Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin B antibody (ab16045)This image is courtesy of an Abreview submitted by Dr Kirk McManusab16045 (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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin B antibody (ab16045)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
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Western blot - Anti-Cyclophilin B antibody (ab16045)Image courtesy of Dr M Schrader by Abreview.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. -
Western blot - Anti-Cyclophilin B antibody (ab16045)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 -
Western blot - Anti-Cyclophilin B antibody (ab16045)
Protocols
References
This product has been referenced in:
- Anderson EM et al. Lactobacillus gasseri CRISPR-Cas9 characterization In Vitro reveals a flexible mode of protospacer-adjacent motif recognition. PLoS One 13:e0192181 (2018). Read more (PubMed: 29394276) »
- Di Pardo A et al. Defective Sphingosine-1-phosphate metabolism is a druggable target in Huntington's disease. Sci Rep 7:5280 (2017). WB . Read more (PubMed: 28706199) »
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