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 conjugated to KLH derived from within residues 150 to the C-terminus of Human Cyclophilin B.

    Read Abcam's proprietary immunogen policy (Peptide available as ab16277.)

  • Positive control
    • This antibody gave a positive signal in: (WB) HeLa; HeLa Nuclear; Jurkat; A431; HEK293; NIH 3T3; Rat Liver Tissue: (IF) NIH3T3.

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

  • 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:
  • 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) »
  • Wang S  et al. The CPLANE protein Intu protects kidneys from ischemia-reperfusion injury by targeting STAT1 for degradation. Nat Commun 9:1234 (2018). Read more (PubMed: 29581513) »
See all 46 Publications for this product

Customer reviews and Q&As

1-10 of 19 Abreviews or Q&A

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

Question
Answer

Vielen Dank für Ihre Anfrage.
Es tut mir leid, dass Sie Probleme mit diesem Antikörper hatten. Wie am Telefon besprochen habe ich eine kostenlose Ersatzlieferung für Sie in Auftrag gegeben. Sie hat die Referenznummer 123456 und sollte morgen bei Ihnen ankommen.
Des Weiteren habe ich noch den Fragebogen angehängt, auf dass wir uns einen Überblick bezüglich des Protokolls machen können.
Ich wünsche Ihnen viel Erfolg für Ihr Projekt. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

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Abcam guarantees this product to work in the species/application used in this Abreview.
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

Question
Answer

Thank you for your phone call.

DISCOUNT CODE: xxx
Expiration date: xxx

I am very pleased to hear you would like to accept our offer and test ab16045 in horse. This code will give you: 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for horse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active.
For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.

Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.

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Answer

Thank you very much for your reply and for sending the images.

Please keep me updated about the results with the replacement vial, and let me know if there is anything else that we can do for you.

Have a nice day.

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Answer

Thank you very much for your call today and for letting us know about the trouble with the new vial of this antibody.

As we discussed, I'm sending a free of charge vial from a new lot on the order ***, which should arrive tomorrow. Please keep me updated about the results with this new vial.

Also, we would be very appreciative if you could please send any images and a detailed protocol (including antibody dilution, blocking solution, etc) so that we can look into the problem with this recent vial.

I look forward to hearing from you. If you have any questions or if there is anything else that we can do for you, please let me know and I'll be happy to help.

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Question
Answer

Thank you for contacting us. I do hope the changes which we have talked about will help with your staining. As you have requested I have attached our ICC protocol which details alternative fixations to PFA. I have also created a 100% Abreview discount code for you. The code and additional information are below. I am very pleased to hear you would like to accept our offer and test ab16045 in Horse. This code will give you: 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for Horse and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you for contacting us.

I have tried to correlate the human housekeeping genes with chicken and then tried to find the whether these are expressed in Chicken heart. The following is the link to human housekeeping genes

http://www.cgen.com/supp_info/housekeeping_genes.html

Beta catenin is protein I have chose which is expressed in Chicken heart, evidence is in publicaiton Fig 5. http://www.plosone.org/article/info:doi%2F10.1371%2Fjournal.pone.0002857#pone-0002857-g004



The anti beta catenin antibodies are ab2365, ab23671,ab16051 and ab79089

Talin 1



Publication; http://www.biomedcentral.com/1471-2121/7/40

ab71333, ab95034

Aspartate aminotransferase



http://pubs.acs.org/doi/abs/10.1021/bi00841a018

ab59132, ab98228

Please note our recommendations are based on the data published in publications and Chicken tested antibodies. It is more likely that the antibodies we have might not have tested in chicken heart however as long as the protein is produced in heart the antibodies will be OK.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Answer

The two lysis buffer recipes in our western blotting guide, one for cytoskeletal protein lysis buffer and one for RIPA, are almost identical except for the addition of EDTA, EGTA, and glycerol in the cytoskeletal protein lysis buffer. I see no reason why the RIPA buffer would not recover as much protein as the cystoskeletal protein lysis buffer from chicken embryo heart.

Have you tried staining a blot of the heart homgenates prepared with the cystoskeletal protein lysis buffer? It is likely that this buffer is as effective as RIPA for recovering cyclophilin from your samples.

I do not think I can explain why the yield of total protein from RIPA is relatively low, if that is in fact the case. The other possibility I mentioned is the effect of the buffer reagents on the BCA assay, though as you have pointed out, they should all be within the acceptable limits of the assay. Are you confident the RIPA buffer was prepared correctly?

One way to check would be to prepare solutions of BSA in each buffer, of equal amounts by weight per volume, for example 1 mg/ml, and then compare the OD in the assay.

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1-10 of 19 Abreviews or Q&A

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