Overview

  • Product name
  • Description
    Rabbit polyclonal to Bestrophin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, WB, ICC, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Human
    Predicted to work with: Cynomolgus monkey
  • Immunogen

    Synthetic peptide:

    LLYGEFLIFLLCYYIIRFIYRL

    , corresponding to amino acids 31-52 of Human Bestrophin. The peptide was amidated for conjugation.

  • Positive control
    • Conditioned medium cultured RPE cells (the basal expression of Bestrophin is low in cultured cells).

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • Storage buffer
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: Tris glycine, 0.5% BSA, 30% Glycerol
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab14927 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
IP
WB
ICC
ICC/IF
  • Application notes
    ICC: Use at an assay dependent dilution.
    IP: 1/200.
    WB: 1/500. Predicted molecular weight: 68 kDa.

    Not tested in other applications.
    Optimal dilutions/concentrations should be determined by the end user.
  • Target

    • Function
      Forms calcium-sensitive chloride channels. Highly permeable to bicarbonate.
    • Tissue specificity
      Predominantly expressed in the basolateral membrane of the retinal pigment epithelium.
    • Involvement in disease
      Defects in BEST1 are the cause of vitelliform macular dystrophy type 2 (VMD2) [MIM:153700]; also known as Best macular dystrophy (BMD). VMD2 is an autosomal dominant form of macular degeneration that usually begins in childhood or adolescence. VMD2 is characterized by typical 'egg-yolk' macular lesions due to abnormal accumulation of lipofuscin within and beneath the retinal pigment epithelium cells. Progression of the disease leads to destruction of the retinal pigment epithelium and vision loss.
      Defects in BEST1 are the cause of retinitis pigmentosa type 50 (RP50) [MIM:613194]. A retinal dystrophy belonging to the group of pigmentary retinopathies. RP is characterized by retinal pigment deposits visible on fundus examination and primary loss of rod photoreceptor cells followed by secondary loss of cone photoreceptors. Patients typically have night vision blindness and loss of midperipheral visual field. As their condition progresses, they lose their far peripheral visual field and eventually central vision as well.
      Defects in BEST1 are a cause of adult-onset vitelliform macular dystrophy (AVMD) [MIM:608161]. AVMD is a rare autosomal dominant disorder with incomplete penetrance and highly variable expression. Patients usually become symptomatic in the fourth or fifth decade of life with a protracted disease of decreased visual acuity.
      Defects in BEST1 are the cause of bestrophinopathy autosomal recessive (ARB) [MIM:611809]. A retinopathy characterized by central visual loss, an absent electro-oculogram light rise, and a reduced electroretinogram.
      Defects in BEST1 are the cause of vitreoretinochoroidopathy autosomal dominant (ADVIRC) [MIM:193220]. A disorder characterized by vitreoretinochoroidal dystrophy. The clinical presentation is variable and may be associated with cataract, nanophthalmos, microcornea, shallow anterior chamber, and glaucoma.
    • Sequence similarities
      Belongs to the bestrophin family.
    • Post-translational
      modifications
      Phosphorylated by PP2A.
    • Cellular localization
      Cell membrane. Basolateral cell membrane.
    • Information by UniProt
    • Database links
    • Alternative names
      • ARB antibody
      • BEST 1 antibody
      • BEST antibody
      • Best disease antibody
      • Best macular dystrophy antibody
      • BEST1 antibody
      • BEST1_HUMAN antibody
      • Best1V1Delta2 antibody
      • Bestrophin 1 antibody
      • Bestrophin-1 antibody
      • Bestrophin1 antibody
      • BMD antibody
      • mBest1 antibody
      • RP50 antibody
      • TU15B antibody
      • Vitelliform macular dystrophy 2 antibody
      • Vitelliform macular dystrophy antibody
      • Vitelliform macular dystrophy protein 2 antibody
      • VMD 2 antibody
      • VMD2 antibody
      see all

    Images

    • ab14927 staining HEK-293 cells transfected with human Bestrophin 1 by ICC/IF.  Cells were PFA fixed and permeabilized in 0.5% Triton X-100 prior to blocking in 5% goat serum for 20 minutes at 25°C.  The primary antibody was diluted 1/100 and incubated with the sample for 12 hours at 4°C.  An Alexa Fluor® 488 conjugated goat anti-rabbit antibody was used as the secondary.  Bestrophin transfected in HEK-293 cells is labeled with green fluorescence. Nuclei were counterstained with DAPI (blue).

      See Abreview

    • All lanes : Anti-Bestrophin antibody (ab14927) at 1/200 dilution

      Lane 1 : Whole cell lysate from bovine retinal pigment epithelial cells
      Lane 2 : Whole cell lysate from non transfected HEK 293 cells

      Lysates/proteins at 25 µg per lane.

      Secondary
      All lanes : HRP conjugated goat anti-rabbit at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 68 kDa
      Observed band size: 68 kDa
      Additional bands at: 74 kDa (possible non-specific binding)


      Exposure time: 20 minutes


      The primary antibody was incubated with the membrane for 12 hours at 4°C.

      See Abreview

    References

    This product has been referenced in:
    • Geng Z  et al. Generation of retinal pigmented epithelium from iPSCs derived from the conjunctiva of donors with and without age related macular degeneration. PLoS One 12:e0173575 (2017). Read more (PubMed: 28282420) »
    • Lin YH  et al. Opening a New Time Window for Treatment of Stroke by Targeting HDAC2. J Neurosci 37:6712-6728 (2017). Read more (PubMed: 28592694) »
    See all 4 Publications for this product

    Customer reviews and Q&As

    1-7 of 7 Abreviews or Q&A

    Answer

    Thank you for your response.

    It is important to clarify that the discount code will be activated after submission of the Abreview and there is normally an expiry date (4 month after offering the code) for sending the Abreview.

    I would advise you to visit the following site where the Terms and Conditions of our testing initiative can be found at: https://www.abcam.com/collaborationdiscount.

    If you need any further assistance in the future, please do not hesitate to contact me.

    Read More

    Answer

    Thank you very much for your interest in ab14927.

    To our knowledge,ab14927 has not been tested in Cell sorting/FACS. Therefore, I can offer a discount off a future purchase if you buyab14927 now, test it inCell sorting/FACS and submit feedback to us in the form of an Abreview. It doesn’t matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of: 1 free PRIMARY ANTIBODY.

    If you are interested in this offer, please follow these steps:

    1. Reply to this e-mail to let me know that you would like to proceed and testab14927 in Cell sorting/FACS. I will then send a discount code. This code must be issued before purchasing abab14927 so please wait for my reply before ordering.

    2. Purchaseab14927 either by phone, fax, or online (www.abcam.com).

    3. Test it in Cell sorting/FACS.

    4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

    5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any PRIMARY ANTIBODY /PROTEIN ordered and the discount code is valid for 4 months after issue.
    Please remember that submission of the Abreview is sufficient for the discount code to become active.

    We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even ifab14927 turns out to be unsuitable for Cell sorting/FACS, you will still receive the discount on your next purchase after your Abreview has been submitted.

    Please let me know if you have any questions about this offer and I would be happy to help you further.

    The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Cow Cell lysate - whole cell (RPE, bovine retinal pigment epithelial cells)
    Loading amount
    25 µg
    Specification
    RPE, bovine retinal pigment epithelial cells
    Gel Running Conditions
    Reduced Denaturing (10%)
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

    Herr Dr. Vladimir Milenkovic

    Verified customer

    Submitted Feb 22 2008

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HEK-293 cells transfected with human Bestrophin 1)
    Specification
    HEK-293 cells transfected with human Bestrophin 1
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% TX 100, 5% goat serum, 1XPBS
    Blocking step
    goat serum as blocking agent for 20 minute(s) · Concentration: 5% · Temperature: 25°C

    Herr Dr. Vladimir Milenkovic

    Verified customer

    Submitted Feb 15 2008

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - whole cell (HEK-293 cells transfected with hBestrophin 1)
    Total protein in input
    20 µg
    Specification
    HEK-293 cells transfected with hBestrophin 1
    Immuno-precipitation step
    Protein G

    Herr Dr. Vladimir Milenkovic

    Verified customer

    Submitted Feb 15 2008

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK 293 transfected with human bestrophin 1)
    Loading amount
    20 µg
    Specification
    HEK 293 transfected with human bestrophin 1
    Gel Running Conditions
    Reduced Denaturing (10 %)
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

    Herr Dr. Vladimir Milenkovic

    Verified customer

    Submitted Feb 15 2008

    Answer

    Thank you for your email. At this time, the blocking peptide is not available for sale. Without knowing additional details regarding your protocol, I would like to make the following suggestions in order to try to help you out. Decrease the amount of primary antibody that you are using (ab14927 is recommended for use at 1:500 in WB, try 1:1000 instead). Decrease the incubation period with the primary -try 2 hrs at RT rather than overnight at 4C. Also, try decreasing the concentration of your secondary antibody as well as the incubation. Make sure to run a secondary-only control to ensure that these bands are not due to the secondary antibody. Make sure that you incorporate sufficient protease inhibitors in your sample buffer, and try boiling in SDS-Page for 10 minutes rather than 5 minutes to disrupt dimers. I hope this helps. Please don't hesitate to contact us again if you need additional assistance.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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