Recombinant
RabMAb

Recombinant Anti-CYP27B1 antibody [EPR20271] (ab206655)

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 within Human CYP27B1 aa 200 to the C-terminus. The exact sequence is proprietary.
    Database link: O15528

  • 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

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

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

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20271
  • Isotype

    IgG

Applications

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.

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.
  • Information by UniProt
  • Database links

  • 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

  • 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.

  • 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.

  • 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).

  • 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).

  • 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.

  • 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.

  • 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.

References

ab206655 has not yet been referenced specifically in any publications.

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