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  1. Link

    cyp27b1-antibody-epr20271-ab206655.pdf

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RecombinantRabMAb

Recombinant Anti-CYP27B1 antibody [EPR20271] (ab206655)

  • Datasheet
  • SDS
  • Certificate of Compliance
Submit a review Submit a question References (6)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP27B1 antibody [EPR20271] (ab206655)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP27B1 antibody [EPR20271] (ab206655)
  • Western blot - Anti-CYP27B1 antibody [EPR20271] (ab206655)
  • Western blot - Anti-CYP27B1 antibody [EPR20271] (ab206655)
  • Western blot - Anti-CYP27B1 antibody [EPR20271] (ab206655)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP27B1 antibody [EPR20271] (ab206655)
  • Immunoprecipitation - Anti-CYP27B1 antibody [EPR20271] (ab206655)
  • 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

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Overview

  • 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.

  • 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.
  • 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
    For more information see here.

    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

  • 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

  • Alternative Versions

    • Anti-CYP27B1 antibody [EPR20271] - BSA and Azide free (ab238978)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)

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).
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.
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

  • 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.
  • Target information above from: UniProt accession O15528 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • Unigene: 6216 Mouse
    • Unigene: 10847 Rat
    see all
  • Alternative names

    • 1alpha(OH)ase antibody
    • 25-hydroxyvitamin D(3) 1-alpha-hydroxylase antibody
    • 25-hydroxyvitamin D-1 alpha hydroxylase antibody
    • 25-OHD-1 alpha-hydroxylase antibody
    • Calcidiol 1-monooxygenase antibody
    • CP27B_HUMAN antibody
    • CP2B antibody
    • CYP1 antibody
    • CYP1ALPHA antibody
    • CYP27B antibody
    • Cyp27b1 antibody
    • Cytochrome p450 27B1 antibody
    • Cytochrome p450 27B13 antibody
    • Cytochrome P450 family 27 subfamily B polypeptide 1 antibody
    • Cytochrome P450 subfamily XXVIIB (25-hydroxyvitamin D-1-alpha-hydroxylase) polypeptide 1 antibody
    • Cytochrome P450 subfamily XXVIIB polypeptide 1 antibody
    • Cytochrome P450C1 alpha antibody
    • Cytochrome P450VD1-alpha antibody
    • mitochondrial antibody
    • P450C1 alpha antibody
    • P450c1 antibody
    • P450C1-alpha antibody
    • P450VD1-alpha antibody
    • PDDR antibody
    • VD3 1A hydroxylase antibody
    • VDD1 antibody
    • VDDR antibody
    • VDDR I antibody
    • VDDRI antibody
    • VDR antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP27B1 antibody [EPR20271] (ab206655)
    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)
    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.

  • Western blot - Anti-CYP27B1 antibody [EPR20271] (ab206655)
    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 minutes


    Blocking/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).

  • Western blot - Anti-CYP27B1 antibody [EPR20271] (ab206655)
    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 seconds


    Blocking/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)
    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 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times: Lane 1, 2, 3 and 4: 1 second; Lane 5: 3 seconds.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CYP27B1 antibody [EPR20271] (ab206655)
    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)
    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.

  • Anti-CYP27B1 antibody [EPR20271] (ab206655)
    Anti-CYP27B1 antibody [EPR20271] (ab206655)

Protocols

  • Western blot protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (6)

Publishing research using ab206655? Please let us know so that we can cite the reference in this datasheet.

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

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