Recombinant Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)
![Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)](https://www.abcam.com/ps/products/178/ab178397/Images/ab178397-412694-anti-cyclophilin-b-antibody-epr12703b-loading-control-western-blot-human-knockout-lysate.jpg "Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR12703(B)] to Cyclophilin B - Loading Control
- Suitable for: WB
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control
See all Cyclophilin B primary antibodies -
Description
Rabbit monoclonal [EPR12703(B)] to Cyclophilin B - Loading Control -
Host species
Rabbit -
Tested applications
Suitable for: WBmore details
Unsuitable for: Flow Cyt,ICC/IF or IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HepG2, HeLa, NIH/3T3, PC-12, HAP1, Jurkat, U87-MG and A431 cell lysates, mouse and rat heart tissue lysates.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR12703(B) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
- HeLa whole cell lysate (ab150035)
- Hep G2 whole cell lysate (ab166833)
- HeLa whole cell lysate (ab29545)
- A-431 whole cell lysate (ab30132)
- Mouse heart normal tissue lysate - total protein (ab30291)
- NIH/3T3 whole cell lysate (ab7179)
- Mouse heart tissue lysate - total protein (0 days) (ab7193)
- Mouse heart tissue lysate - total protein (14 days) (ab7194)
- A-431 whole cell lysate (ab7909)
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab178397 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 | (4) |
1/1000 - 1/10000. Predicted molecular weight: 24 kDa.
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| Notes |
|---|
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WB
1/1000 - 1/10000. Predicted molecular weight: 24 kDa. |
Target
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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
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Database links
- Entrez Gene: 5479 Human
- Entrez Gene: 19035 Mouse
- Entrez Gene: 64367 Rat
- Omim: 123841 Human
- SwissProt: P23284 Human
- SwissProt: P24369 Mouse
- SwissProt: P24368 Rat
- Unigene: 434937 Human
see all -
Alternative names
- AA408962 antibody
- AA553318 antibody
- AI844835 antibody
see all
Images
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Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)All lanes : Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397) at 1/1000 dilution
Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : PPIB knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 24 kDa
Observed band size: 24 kDaLanes 1-4: Merged signal (red and green). Green - ab178397 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa.
ab178397 Anti-Cyclophilin B antibody [EPR12703(B)] was shown to specifically react with Cyclophilin B in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261746 (knockout cell lysate ab257037) was used. Wild-type and Cyclophilin B knockout samples were subjected to SDS-PAGE. ab178397 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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![Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)](https://www.abcam.com/ps/products/178/ab178397/Images/ab178397-349450-anti-cyclophilin-b-antibody-epr12703b-loading-control-western-blot-knock-out-hap1-ppib-jurkat-u87-mg-human-lysates.jpg)
Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)All lanes : Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397) at 1 µg/ml
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: 24 kDaLanes 1 - 4: Merged signal (red and green). Green - ab178397 observed at 24 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab178397 was shown to recognize PPIB in wild-type HAP1 cells as signal was lost at the expected MW in PPIB knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and PPIB knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab178397 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml 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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![Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)](https://www.abcam.com/ps/products/178/ab178397/Images/ab178397-253079-anti-cyclophilin-b-antibody-epr12703b-loading-control-western-blot.jpg)
Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)All lanes : Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397) at 1/10000 dilution (purified)
Lane 1 : Mouse heart tissue lysate
Lane 2 : NIH/3T3 cell lysate
Lane 3 : Rat heart tissue lysate
Lane 4 : PC-12 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution
Predicted band size: 24 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
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![Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)](https://www.abcam.com/ps/products/178/ab178397/Images/ab178397-253078-anti-cyclophilin-b-antibody-epr12703b-loading-control-western-blot.jpg)
Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)All lanes : Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397) at 1/10000 dilution (purified)
Lane 1 : HepG2 cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : A431 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution
Predicted band size: 24 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
-
![Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)](https://www.abcam.com/ps/products/178/ab178397/Images/ab178397-206289-anti-cyclophilin-b-antibody-epr12703b-loading-control-western-blot.jpg)
Western blot - Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)All lanes : Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397) at 1/1000 dilution
Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 3 : A431 (Human epidermoid carcinoma cell line) cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Developed using the ECL technique.
Predicted band size: 24 kDa -
![Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)](https://www.abcam.com/ps/products/178/ab178397/Images/ab178397-6-benefits-of-recombinant-antibodies.png)
Anti-Cyclophilin B antibody [EPR12703(B)] - Loading Control (ab178397)
Datasheets and documents
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SDS download
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Datasheet download
References (14)
ab178397 has been referenced in 14 publications.
- Navarro-Guerrero E et al. Genome-wide CRISPR/Cas9-knockout in human induced Pluripotent Stem Cell (iPSC)-derived macrophages. Sci Rep 11:4245 (2021). PubMed: 33608581
- Kim J et al. The hexosamine biosynthesis pathway is a targetable liability in KRAS/LKB1 mutant lung cancer. Nat Metab 2:1401-1412 (2020). PubMed: 33257855
- Buel GR et al. Structure of E3 ligase E6AP with a proteasome-binding site provided by substrate receptor hRpn10. Nat Commun 11:1291 (2020). PubMed: 32157086
- Nagaraja S et al. Histone Variant and Cell Context Determine H3K27M Reprogramming of the Enhancer Landscape and Oncogenic State. Mol Cell 76:965-980.e12 (2019). PubMed: 31588023
- Lee E et al. GREB1 amplifies androgen receptor output in human prostate cancer and contributes to antiandrogen resistance. Elife 8:N/A (2019). PubMed: 30644358