Anti-Cyclophilin B 抗体 (ab16045)
製品の概要
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製品名
Anti-Cyclophilin B antibody
Cyclophilin B 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to Cyclophilin B -
由来種
Rabbit -
アプリケーション
適用あり: WB, IHC-P, ICC/IF, IHC-Fr, IPmore details -
種交差性
交差種: Mouse, Rat, Horse, Chicken, Dog, Human
交差が予測される動物種: Cow, Pig, Xenopus laevis
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免疫原
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ポジティブ･コントロール
- WB: HeLa, HeLa Nuclear, Jurkat, A431, HEK293, NIH 3T3, HAP1 and U87-MG cell lysates; Rat Liver Tissue IF: NIH3T3 cells
製品の特性
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製品の状態
Liquid -
保存方法
Shipped 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. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading... -
精製度
Immunogen affinity purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Positive Controls
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Recombinant Protein
アプリケーション
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.
| アプリケーション | Abreviews | 特記事項 |
|---|---|---|
| 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. |
ターゲット情報
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機能
PPIases accelerate the folding of proteins. It catalyzes the cis-trans isomerization of proline imidic peptide bonds in oligopeptides. -
関連疾患
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. -
配列類似性
Belongs to the cyclophilin-type PPIase family. PPIase B subfamily.
Contains 1 PPIase cyclophilin-type domain. -
細胞内局在
Endoplasmic reticulum lumen. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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参照データベース
- Entrez Gene: 5479 Human
- Entrez Gene: 19035 Mouse
- Entrez Gene: 64367 Rat
- Omim: 123841 Human
- SwissProt: P24367 Chicken
- SwissProt: P80311 Cow
- SwissProt: P23284 Human
- SwissProt: P24369 Mouse
see all -
別名
- AA408962 antibody
- AA553318 antibody
- AI844835 antibody
see all
画像
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Western blot - Anti-Cyclophilin B antibody (ab16045)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.
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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
プロトコール
データシートおよび資料
参考文献
ab16045 は 60 報の論文で使用されています。
- Chavez RD et al. Prg4 prevents osteoarthritis induced by dominant-negative interference of TGF-ß signaling in mice. PLoS One 14:e0210601 (2019). 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). PubMed: 30630053
- Mumin NH et al. Overcoming acquired resistance to HSP90 inhibition by targeting JAK-STAT signalling in triple-negative breast cancer. BMC Cancer 19:102 (2019). PubMed: 30678647
- Di Pardo A et al. Stimulation of Sphingosine Kinase 1 (SPHK1) Is Beneficial in a Huntington's Disease Pre-clinical Model. Front Mol Neurosci 12:100 (2019). PubMed: 31068790
- Jo S et al. Type 2 deiodinase polymorphism causes ER stress and hypothyroidism in the brain. J Clin Invest 129:230-245 (2019). PubMed: 30352046
