PHOSPHO1 Antibody | 5643

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PHOSPHO1 Antibody | 5643 gallery image 1

Western blot analysis of HEK293 human embryonic kidney whole cell lysate probed with Human anti Human PHOSPHO1 antibody (HCA093) followed by HRP conjugated F(ab')2 Goat anti Human IgG, visualized by chemiluminescence

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Published customer image:
Human anti PHOSPHO1 antibody, clone 5643 used for the evaluation of PHOSPHO1 expression in MC3T3-C14 cells in vitro by western blotting.
Image caption:
Extracellular matrix mineralisation and temporal gene expression in MC3T3-C14 cells. a Alizarin red staining and b relative quantification of cetylpyridinium chloride leached bound dye. RT-qPCR analysis of cPhospho1dAlpl and eSmpd3 mRNA expression in mineralising MC3T3-C14 cells over a 10-day culture period. f Western blotting analysis of PHOSPHO1, TNAP and nSMase2 in MC3T3-C14 cells over a 10-day culture period. The fold change in fluorescence intensity against cultures at day 0 (normalised against beta-actin) is shown below each protein of interest. g Assessment of matrix mineralisation in MC3T3-C14 cell cultures continuously exposed to PTH for 10 days. Quantification of, cetylpyridinium chloride leached, bound Alizarin red staining. N = 3; *P <0.05, **P <0.01; ***P <0.001 in comparison with the previous time point in culture.

From: Houston DA, Myers K, MacRae VE, Staines KA, Farquharson C.
The Expression of PHOSPHO1, nSMase2 and TNAP is Coordinately Regulated by Continuous PTH Exposure in Mineralising Osteoblast Cultures.
Calcif Tissue Int. 2016 Nov;99(5):510-524.

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PHOSPHO1 Antibody | 5643 gallery image 3

Published customer image:
Human anti PHOSPHO1 antibody, clone 5643 used for the evaluation of PHOSPHO1 expression in MC3T3-C14 cells in vitro by western blotting.
Image caption:
Regulation of key mineralisation genes by PTH in MC3T3-C14 cells. RT-qPCR analysis of aPhospho1, bAlpl and cSmpd3 mRNA expression in response to a 24-h exposure of various doses of bPTH (1–34) on day 6 of culture. RT-qPCR analysis of dPhospho1, eAlpl and fSmpd3 mRNA expression in response to various exposure times of bPTH (1–34) (50 nM). The timing of PTH addition was adjusted to ensure all experiments finished on day 7 of culture. RT-qPCR analysis of gPhospho1, hAlpl and iSmpd3 mRNA expression in response to 24-h exposure PTH (50 nM) in MC3T3-C14 cells at different stages of differentiation (confluency (day 0), days 7 and day 10 post-confluency). j Western blotting analysis of PHOSPHO1, TNAP, nSMase2 in response to a 24-h exposure of various doses of PTH. The fold change in fluorescence intensity against control treated cultures (normalised against beta-actin) is shown below each protein of interest. k Western blotting analysis of PHOSPHO1, TNAP, nSmase2 in response to various exposure times of PTH (50 nM). The fold change in fluorescence intensity against control treated cultures (normalised against beta-actin) is shown below each protein of interest. l Western blotting analysis of PHOSPHO1, TNAP, nSMase2 in response a 24-h exposure PTH (50 nM) in MC3T3-C14 cells at different stages of differentiation. The fold change in fluorescence intensity against day 0 control treated cultures (normalised against beta-actin) is shown below each protein of interest. N = 3; *P <0.05; **P <0.01; ***P <0.001 in comparison with controls.

From: Houston DA, Myers K, MacRae VE, Staines KA, Farquharson C.
The Expression of PHOSPHO1, nSMase2 and TNAP is Coordinately Regulated by Continuous PTH Exposure in Mineralising Osteoblast Cultures.
Calcif Tissue Int. 2016 Nov;99(5):510-524.

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  • Human anti PHOSPHO1
(Rated 0.0 out of 5 based on 0 customer reviews)
  • Product Type
    Monoclonal Antibody
  • Clone
    5643
  • Isotype
    HuCAL Fab bivalent
1 Formats Available
    Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
    HCA093E, WBdatasheet pdfdatasheet pdf0.1 mg
    HCA093
    Summary
    Secondary Antibodies
    Negative Isotype Controls
    Useful Reagents
    Positive Controls
    Histology Controls
    More Images
    References
    Reviews
    -
    • Human anti PHOSPHO1, clone 5643 recognizes human phosphoethanolamine/phosphocholine phosphatase (PHOSPHO1), a phosphatase that belongs to the haloacid dehalogenase (HAD) superfamily of hydrolases. It is highly expressed in skeletal tissues, expression being localised to sites of mineralization in cartilage and bone. Reports suggest that PHOSPHO1 may be involved in the mineralization process in both osteoblasts and chrondrocytes.
    • Intended Use
    • Target Species
      Human
    • Species Cross-Reactivity
      Target SpeciesCross Reactivity
      Mouseyes
      N.B. Antibody reactivity and working conditions may vary between species.
    • Product Form
      A bivalent human recombinant Fab selected from the HuCAL® phage display library, expressed in E. coli. The antibody is tagged with a myc-tag (EQKLISEEDL) and a his-tag (HHHHHH) at the C-terminus of the antibody heavy chain. This Fab fragment is dimerized via a helix-turn-helix motif. This antibody is supplied as a liquid.
    • Reconstitution
    • Preparation
      Metal chelate affinity chromatography
    • Preservative Stabilisers
      0.01% Thiomersal
    • Immunogen
      Purified recombinant human PHOSPHO1
    • Purity
    • Approx. Protein Concentrations
      Total protein concentration 0.5mg/ml
    • Reagents In The Kit
    • Preparing The Antibody
    • Test Principle
    • Buffer Solution
      Phosphate buffered saline
    • Storage
      Store at +4oC or at -20oC if preferred.
      Storage in frost-free freezers is not recommended.
      This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use.
    • Shelf Life
      12 months from date of despatch.
    • GO Terms
      metal ion binding
      regulation of bone mineralization
      phosphoethanolamine/phosphocholine phosphatase activity
    • UniProt
    • Entrez Gene
    • Acknowledgements
      Sold under license of U.S. Patents 6,300,064, 6,696,248, 6,708,484, 6,753,136, European patent 0,859,841 and corresponding patents.
    • Licensed Use
      For in vitro research purposes only, unless otherwise specified in writing by Bio-Rad.
    • Regulatory
      For research purposes only
    • This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.

    • Application NameYesNoMin DilutionMax Dilution
      ELISA1/5001/5000
      Western Blotting

    • Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
    • Technical Advice
      Recommended protocols and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual
    • Recommended Protocol
    • ELISA
    • Immunohistology
    • Histology Positive Control Tissue
    • Immunofluorescence
    • Western Blotting
      HCA093 detects a band of approximately 32kDa in mouse bone extracts
    • Instructions For Use

    Additional PHOSPHO1 Antibody Formats

    Formats Clone Applications Sizes available
    PHOSPHO1 Antibody : Purified 5643 E, WB 0.1 mg
    • Copyright © 2016 Bio-Rad Antibodies (formerly AbD Serotec)

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      Recommended Negative Isotype Control

        Useful Reagents

          Recommended Positive Controls

            Histology Controls

              • Application NameReference Images
                Western Blotting

              References

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