Anti-Cyclophilin B抗体
参阅全部 Cyclophilin B 一抗
兔多克隆抗体to Cyclophilin B
Rabbit
Replenishment batches of our polyclonal antibody, ab16045 are tested in WB. Previous batches were additionally validated in ICC/IF and IP. These applications are still expected to work and are covered by our Abpromise guarantee. You may also be interested in our alternative recombinant antibody, ab178397.
适用于: WB, ICC/IF, IPmore details
与反应: Mouse, Rat, Horse, Chicken, Dog, Human
预测可用于: Cow, Pig, Xenopus laevis
Synthetic peptide corresponding to Human Cyclophilin B aa 150 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
(Peptide available as ab16277, ab5016)
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Liquid
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.
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA
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.
浓度
批次浓度范围 100 µg 浓度为 0.8 - 1 mg/ml
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab16045于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB | (7) | Use a concentration of 0.5 µg/ml. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa). |
| ICC/IF | (3) | Use a concentration of 1 µg/ml. |
| IP | Use at an assay dependent concentration. |
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Unigene: 434937 Human
Unigene: 335249 Mouse
Unigene: 1893 Rat
Entrez Gene: 5479 Human
Entrez Gene: 19035 Mouse
Omim: 123841 Human
SwissProt: P24367 Chicken
SwissProt: P23284 Human
SwissProt: P24369 Mouse
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Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin B antibody (ab16045)
ab16045 staining Cyclophilin B in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab16045 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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 kDawhy 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.
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 kDawhy 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 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
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).
Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin B antibody (ab16045)This image is courtesy of an Abreview submitted by Dr Kirk McManus
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.
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 kDawhy 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
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