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INPP5E antibody (C-Term)

INPP5E Reactivity: Human, Mouse, Rat WB Host: Rabbit Polyclonal unconjugated
Catalog No. ABIN7298962
  • Target See all INPP5E Antibodies
    INPP5E (Inositol Polyphosphate-5-Phosphatase, 72 KDa (INPP5E))
    Binding Specificity
    • 4
    • 3
    • 1
    • 1
    C-Term
    Reactivity
    Human, Mouse, Rat
    Host
    • 16
    • 2
    Rabbit
    Clonality
    • 18
    Polyclonal
    Conjugate
    • 8
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    This INPP5E antibody is un-conjugated
    Application
    • 7
    • 6
    • 4
    • 3
    • 1
    Western Blotting (WB)
    Specificity
    Recognizes endogenous levels of INPP5E protein.
    Characteristics
    Rabbit polyclonal antibody to INPP5E
    Purification
    The antibody was purified by immunogen affinity chromatography.
    Immunogen
    KLH-conjugated synthetic peptide encompassing a sequence within the C-term region of human INPP5E.
  • Application Notes
    WB (1:500 - 1:1000)
    Restrictions
    For Research Use only
  • Validation #101129 (Western Blotting)
    'Independent Validation' Badge
    by
    Children's Hospital of Eastern Ontario Research Institute
    No.
    #101129
    Date
    11/10/2017
    Antigen
    INPP5E
    Lot Number
    CP1D10A
    Method validated
    Western Blotting
    Positive Control
    Inpp5e+/+ 4T1 mouse breast carcinoma cell line (transfected with non-targeting Cas9 gRNA), treated with an inducer (unpublished)
    Negative Control
    Inpp5eem 4T1 cell line (CRISPR/Cas9 endonuclease mediated Inpp5e knock-out)
    Notes

    Passed. ABIN2704925 specifically recognizes Inpp5e in murine Inpp5e+/+ 4T1 cells upon induction aside from other unspecific bands.

    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN2704925
    Secondary Antibody
    goat anti-rabbit IgG (H+L)-HRP-conjugated antibody (BioRad, 172-1019, lot 350001898)
    Full Protocol
    • Grow Inpp5e+/+ and Inpp5eem 4T1 murine cells in HyClone TM RPMI-1640 medium (GE Healthcare Life Science, SH30027.01, lot AC11016269) supplemented with fetal bovine serum (Sigma, F1051-500ML, lot 17D192) and HyClone penicillin – streptomycine solution (GE Healthcare Life Science, SV30010, lot SH40003.01), at 37°C and 5% CO2 in 3ml on a 6-well plate to 95% confluency.
    • Treat cells with an “inducer” (unpublished) to increase Inpp5e expression.
    • Lyse cells in 200µl per well of cold RIPA buffer (150mM NaCl, 1.0% IGEPAL CA-630 Sigma, 0.5% sodium deoxycholate, 0.1% SDS, 50mM Tris, pH8.0).
    • Determine the total protein content of the lysates using DC Protein Assay (BioRad, 500-0114, lot 200008008).
    • Denature 30µg of total protein for 5min at 95°C in 5x Laemmli SDS sample buffer and subsequently separate proteins on a non-/denaturing 10% SDS-polyacrylamide gel in a mini-Protean electrophoresis cell (BioRad) for 90min at 100V.
    • Transfer proteins onto 0.2µm nitrocellulose membrane (BioRad, 1620112, lot A10088580) with a MiniTransblot Cell (BioRad, 1703930) system for 60V at 300mA.
    • Block the membrane with TBS containing 0.1% Tween 20 (TBST) and 5% skim milk for 30min at RT.
    • Incubation with primary rabbit anti-INNP5E antibody (antibodies-online, ABIN2704925, CP1D10A) diluted 1:1000 in TBST containing 5% BSA and 0.05% NaN3 ON at 4°C.
    • Wash membrane 3x with TBST.
    • Incubation with secondary goat anti-rabbit IgG (H+L)-HRP-conjugated antibody (BioRad, 172-1019, lot 350001898) diluted 1:5000 in TBST for 1h at RT.
    • Wash membrane 5x with TBST.
    • Reveal protein bands using Clarity Western ECL Substrate (BioRad, 1705060, lot 102030837) on a ChemiDoc Imaging System (Biorad, 17001401).
    Experimental Notes
    • ABIN2704925 reveals a single band in the "wild type" cell line with treatment. This protein band is abrogated in the CRISPR/Cas9 targeted cells and under non-inducing conditions, thus demonstrating immunospecificity of ABIN2704925 for Inpp5e in the mouse.

    • We observed a lower molecular weight than the predicted size of 72kDa for Inpp5e. We suspect that this might be a splice isoform. We observed a similar band also with a different anti-INPP5E antibody (Thermo Fisher Scientific, PA5-37119) using similar experiment condition.

    • ABIN2704925 also recognizes potentially unspecific bands at various sizes.
  • Format
    Liquid
    Buffer
    Liquid in 0.42 % Potassium phosphate, 0.87 % Sodium chloride, pH 7.3, 30 % glycerol, and 0.01 % sodium azide.
    Preservative
    Sodium azide
    Precaution of Use
    This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Storage
    -20 °C
    Storage Comment
    Shipped at 4°C. Upon delivery aliquot and store at -20°C for one year. Avoid freeze/thaw cycles.
    Expiry Date
    12 months
  • Target
    INPP5E (Inositol Polyphosphate-5-Phosphatase, 72 KDa (INPP5E))
    Alternative Name
    INPP5E (INPP5E Products)
    Synonyms
    1200002L24Rik antibody, 72kDa antibody, mKIAA0123 antibody, CORS1 antibody, CPD4 antibody, JBTS1 antibody, MORMS antibody, PPI5PIV antibody, PMPCA antibody, inositol polyphosphate-5-phosphatase E antibody, Inpp5e antibody, INPP5E antibody
    Background
    72 kDa inositol polyphosphate 5-phosphatase, Phosphatidylinositol 4,5-bisphosphate 5-phosphatase, Phosphatidylinositol polyphosphate 5-phosphatase type IV
    Gene ID
    56623, 64436
    UniProt
    Q9NRR6, Q9JII1, Q9WVR1
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