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Product name
Anti-Cyclophilin B antibody
See all Cyclophilin B products (3) ...
Description
Rabbit polyclonal to Cyclophilin B
Tested applications
WB, IHC-P, ICC/IF, IHC-Fr, IPmore details
Cross reactivity
Reacts with
Mouse, Rat, Chicken, Dog, Human
Predicted to work with
Cow, Pig, Xenopus laevis
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 150 to the C-terminus of Human Cyclophilin B.
(Peptide available as ab162 77.)
Positive control
This antibody gave a positive signal in: (WB) HeLa; HeLa Nuclear; Jurkat; A431; HEK293; NIH 3T3; Rat Liver Tissue: (IF) NIH3T3.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Signal Transduction >> Protein Trafficking >> ER Proteins
Immunology >> Immune Interventions >> Transplantation
Applications
Show applications key
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.
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 Cyclophilin B peptide (ab16277).
IHC-P: Use at an assay dependent dilution. 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.
Target information above from: UniProt accessionP23284
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- AA553318 antibody
- AI844835 antibody
- Cphn 2 antibody
see all
Database links
- Entrez Gene: 281419 Cow
- Entrez Gene: 5479 Human
- Entrez Gene: 19035 Mouse
- Entrez Gene: 64367 Rat
- Omim: 123841 Human
- SwissProt: P24367 Chicken
- SwissProt: P80311 Cow
- SwissProt: Q5E9F4 Cow
see all
Anti-Cyclophilin B antibody images:
Western blot - 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
Goat polyclonal Secondary Antibody to 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
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 : Jukat whole cell lysate
Lane 5 : HEK293 whole cell lysate
Lane 6 : HeLa nuclear lysate with
Lane 7 : HeLa whole cell lysate with
Lane 8 : A431 whole cell lysate with
Lane 9 : Jurkat whole cell lysate with
Lane 10 : HEK293 whole cell lysate with
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal Secondary Antibody to 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 - 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).
Immunocytochemistry/ Immunofluorescence - Cyclophilin B antibody (ab16045)
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.
This image is courtesy of an Abreview submitted by Dr Kirk McManus
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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
.Western blot - Cyclophilin B antibody (ab16045)
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
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.
Image courtesy of Dr M Schrader by Abreview.
Western blot - Anti-Cyclophilin B antibody (ab16045)
Anti-Cyclophilin B antibody (ab16045) at 0.5 µg/ml +
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Exposure time : 30 seconds
References for Anti-Cyclophilin B antibody (ab16045)
This product has been referenced in:
- Sebollela A et al. Amyloid-ß Oligomers Induce Differential Gene Expression in Adult Human Brain Slices. J Biol Chem 287:7436-45 (2012). WB; Rat.Read more (PubMed: 22235132) »
- Wolff NC et al. Cell-type-dependent regulation of mTORC1 by REDD1 and the tumor suppressors TSC1/TSC2 and LKB1 in response to hypoxia. Mol Cell Biol 31:1870-84 (2011).Read more (PubMed: 21383064) »
See all 17 publications for this product
Publishing research using ab16045? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"