Anti-Cyclophilin B抗体(ab16045)

概述

  • 产品名称Anti-Cyclophilin B抗体
    参阅全部 Cyclophilin B 一抗
  • 描述
    兔多克隆抗体to Cyclophilin B
  • 经测试应用适用于: WB, IHC-P, ICC/IF, IHC-Fr, IPmore details
  • 种属反应性
    与反应: Mouse, Rat, Horse, Chicken, Dog, Human
    预测可用于: Cow, Pig, Xenopus laevis
  • 免疫原

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

    (Peptide available as ab16277.)

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

性能

应用

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.

应用 Ab评论 说明
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.

靶标

  • 功能PPIases accelerate the folding of proteins. It catalyzes the cis-trans isomerization of proline imidic peptide bonds in oligopeptides.
  • 疾病相关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.
  • 序列相似性Belongs to the cyclophilin-type PPIase family. PPIase B subfamily.
    Contains 1 PPIase cyclophilin-type domain.
  • 细胞定位Endoplasmic reticulum lumen. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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 图像

  • 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
    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


  • Performed under reducing conditions.

    Predicted band size : 21 kDa
  • 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
    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

    Image courtesy of Dr M Schrader by Abreview.

    See Abreview


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 21 kDa

Anti-Cyclophilin B antibody (ab16045)参考文献

This product has been referenced in:

See all 21 Publications for this product

Product Wall

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
Username

Mr. Jeremy Gingrich

Verified customer

提交于 Oct 30 2015

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 I...

Read More
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
Username

Dr. Shawna Pyott

Verified customer

提交于 Aug 24 2012

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 expirat...

Read More

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.

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 kee...

Read More

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 discou...

Read More

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://...

Read More

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 w...

Read More

Thank you for contacting us.

Can you please tell me what your protein yield was with the cytoskeletal buffer? I am not sure why the RIPA yield would be much lower, unless your protein assay is incompatible with the RIPA buffer components. Ca...

Read More

1-10 of 19 Abreviews or Q&A

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