Recombinant Anti-CYP27B1 antibody [EPR20271] (ab206655)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP27B1 antibody [EPR20271] (ab206655)](https://www.abcam.com/ps/products/206/ab206655/Images/ab206655-274877-anti-cyp27b1-antibody-epr20271-immunohistochemistry.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP27B1 antibody [EPR20271] (ab206655)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20271] to CYP27B1
- Suitable for: WB, IHC-P, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-CYP27B1 antibody [EPR20271]
See all CYP27B1 primary antibodies -
Description
Rabbit monoclonal [EPR20271] to CYP27B1 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HaCaT, MeWo and HeLa whole cell lysates; Human fetal heart, fetal brain, fetal kidney and fetal liver lysates; Mouse heart and kidney lysates; Rat heart, kidney and brain lysates. IHC-P: Human, mouse and rat kidney tissues. IP: MeWo whole cell lysate.
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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.
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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20271 -
Isotype
IgG
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab206655 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 |
1/1000. Detects a band of approximately 45, 70 kDa (predicted molecular weight: 57 kDa).
|
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| IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
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| IP |
1/40.
|
| Notes |
|---|
|
WB
1/1000. Detects a band of approximately 45, 70 kDa (predicted molecular weight: 57 kDa). |
|
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
IP
1/40. |
Target
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Function
Catalyzes the conversion of 25-hydroxyvitamin D3 (25(OH)D) to 1-alpha,25-dihydroxyvitamin D3 (1,25(OH)2D) plays an important role in normal bone growth, calcium metabolism, and tissue differentiation. -
Tissue specificity
Kidney. -
Pathway
Hormone biosynthesis; cholecalciferol biosynthesis. -
Involvement in disease
Defects in CYP27B1 are the cause of rickets vitamin D-dependent type 1A (VDDR1A) [MIM:264700]; also known as pseudovitamin D deficiency rickets (PDDR). A disorder caused by a selective deficiency of the active form of vitamin D (1,25-dihydroxyvitamin D3) and resulting in defective bone mineralization and clinical features of rickets. -
Sequence similarities
Belongs to the cytochrome P450 family. -
Cellular localization
Mitochondrion membrane. - Information by UniProt
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Database links
- Entrez Gene: 1594 Human
- Entrez Gene: 13115 Mouse
- Entrez Gene: 114700 Rat
- Omim: 609506 Human
- SwissProt: O15528 Human
- SwissProt: O35084 Mouse
- SwissProt: O35132 Rat
- Unigene: 524528 Human
see all -
Alternative names
- 1alpha(OH)ase antibody
- 25-hydroxyvitamin D(3) 1-alpha-hydroxylase antibody
- 25-hydroxyvitamin D-1 alpha hydroxylase antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP27B1 antibody [EPR20271] (ab206655)
Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling CYP27B1 with ab206655 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Strong cytoplasmic staining on distal convoluted tubules of human kidney. Weaker cytoplasmic staining on proximal convoluted tubules [PMID: 23416105] [PMID: 12050281].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP27B1 antibody [EPR20271] (ab206655)](https://www.abcam.com/ps/products/206/ab206655/Images/ab206655-274876-anti-cyp27b1-antibody-epr20271-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP27B1 antibody [EPR20271] (ab206655)Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling CYP27B1 with ab206655 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on mouse kidney tubules [PMID: 23416105] [PMID: 12050281].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Western blot - Anti-CYP27B1 antibody [EPR20271] (ab206655)](https://www.abcam.com/ps/products/206/ab206655/Images/ab206655-274880-anti-cyp27b1-antibody-epr20271-western-blot.jpg)
Western blot - Anti-CYP27B1 antibody [EPR20271] (ab206655)All lanes : Anti-CYP27B1 antibody [EPR20271] (ab206655) at 1/2000 dilution
Lane 1 : THP-1 (Human monocytic leukemia cell line) whole cell lysate
Lane 2 : HaCaT (Human keratinocyte cell line) whole cell lysate
Lane 3 : MeWo (Human malignant melanoma cell line) whole cell lysate
Lane 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 5 : Human fetal heart lysate
Lane 6 : Human fetal brain lysate
Lysates/proteins at 20 µg per lane.
Secondary
Lanes 1-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lanes 5-6 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
Predicted band size: 57 kDa
Observed band size: 45,70 kDa why is the actual band size different from the predicted?
Exposure time: 3 minutesBlocking/Dilution buffer: 5% NFDM/TBST.
THP-1 is a negative control (PMID: 16930540).
The molecular weight observed is consistent with what has been described in the literature (PMID: 16061850).
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![Western blot - Anti-CYP27B1 antibody [EPR20271] (ab206655)](https://www.abcam.com/ps/products/206/ab206655/Images/ab206655-274879-anti-cyp27b1-antibody-epr20271-western-blot.jpg)
Western blot - Anti-CYP27B1 antibody [EPR20271] (ab206655)All lanes : Anti-CYP27B1 antibody [EPR20271] (ab206655) at 1/1000 dilution
Lane 1 : Human fetal kidney lysate
Lane 2 : Human fetal liver lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
Predicted band size: 57 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Exposure time: 30 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 16061850).
-
![Western blot - Anti-CYP27B1 antibody [EPR20271] (ab206655)](https://www.abcam.com/ps/products/206/ab206655/Images/ab206655-274878-anti-cyp27b1-antibody-epr20271-western-blot.jpg)
Western blot - Anti-CYP27B1 antibody [EPR20271] (ab206655)All lanes : Anti-CYP27B1 antibody [EPR20271] (ab206655) at 1/1000 dilution
Lane 1 : Mouse heart lysate
Lane 2 : Mouse kidney lysate
Lane 3 : Rat heart lysate
Lane 4 : Rat kidney lysate
Lane 5 : Rat brain lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 57 kDa
Observed band size: 57 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure times: Lane 1, 2, 3 and 4: 1 second; Lane 5: 3 seconds.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP27B1 antibody [EPR20271] (ab206655)](https://www.abcam.com/ps/products/206/ab206655/Images/ab206655-274875-anti-cyp27b1-antibody-epr20271-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP27B1 antibody [EPR20271] (ab206655)Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling CYP27B1 with ab206655 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on rat kidney tubules [PMID: 23416105] [PMID: 12050281].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunoprecipitation - Anti-CYP27B1 antibody [EPR20271] (ab206655)](https://www.abcam.com/ps/products/206/ab206655/Images/ab206655-274874-anti-cyp27b1-antibody-epr20271-immunoprecipitation.jpg)
Immunoprecipitation - Anti-CYP27B1 antibody [EPR20271] (ab206655)CYP27B1 was immunoprecipitated from 0.35 mg of MeWo (Human malignant melanoma cell line) whole cell lysate with ab206655 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab206655 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: MeWo whole cell lysate, 10 μg (Input).
Lane 2: ab206655 IP in MeWo whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab206655 in MeWo whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
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![Anti-CYP27B1 antibody [EPR20271] (ab206655)](https://www.abcam.com/ps/products/206/ab206655/Images/ab206655-8-benefits-of-recombinant-antibodies.png)
Anti-CYP27B1 antibody [EPR20271] (ab206655)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (6)
ab206655 has been referenced in 6 publications.
- Debruin DA et al. Exercise May Ameliorate the Detrimental Side Effects of High Vitamin D Supplementation on Muscle Function in Mice. J Bone Miner Res 35:1092-1106 (2020). PubMed: 32078180
- Mori R et al. Modified expression of vitamin D receptor and CYP27B1 in denervation-induced muscle atrophy. Biochem Biophys Res Commun 529:733-739 (2020). PubMed: 32736700
- Huo X et al. CYP27B1 Downregulation: A New Molecular Mechanism Regulating EZH2 in Ovarian Cancer Tumorigenicity. Front Cell Dev Biol 8:561804 (2020). PubMed: 33163485
- Anisiewicz A et al. Vitamin D Metabolite Profile in Cholecalciferol- or Calcitriol-Supplemented Healthy and Mammary Gland Tumor-Bearing Mice. Nutrients 12:N/A (2020). PubMed: 33172201
- Anisiewicz A et al. Calcitriol in the Presence of Conditioned Media from Metastatic Breast Cancer Cells Enhances Ex Vivo Polarization of M2 Alternative Murine Bone Marrow-Derived Macrophages. Cancers (Basel) 12:N/A (2020). PubMed: 33238581
- Ide N et al. In vivo evidence for an interplay of FGF23/Klotho/PTH axis on the phosphate handling in renal proximal tubules. Am J Physiol Renal Physiol 315:F1261-F1270 (2018). PubMed: 29993278