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SPNS2 antibody (N-Term)

SPNS2 Reactivity: Human, Mouse, Rat, Pig, Rabbit WB Host: Rabbit Polyclonal unconjugated
Catalog No. ABIN2786494
  • Target See all SPNS2 Antibodies
    SPNS2 (Spinster Homolog 2 (SPNS2))
    Binding Specificity
    • 8
    • 7
    • 6
    • 5
    • 3
    • 1
    N-Term
    Reactivity
    • 24
    • 8
    • 4
    • 3
    • 3
    • 1
    • 1
    Human, Mouse, Rat, Pig, Rabbit
    Host
    • 27
    Rabbit
    Clonality
    • 27
    Polyclonal
    Conjugate
    • 11
    • 5
    • 4
    • 3
    • 2
    • 2
    This SPNS2 antibody is un-conjugated
    Application
    • 22
    • 21
    • 4
    • 3
    • 2
    • 1
    • 1
    • 1
    Western Blotting (WB)
    Sequence
    PPGTPGTPGC AATAKGPGAQ QPKPASLGRG RGAAAAILSL GNVLNYLDRY
    Predicted Reactivity
    Human: 100%, Mouse: 85%, Pig: 100%, Rabbit: 92%, Rat: 93%
    Characteristics
    This is a rabbit polyclonal antibody against SPNS2. It was validated on Western Blot using a cell lysate as a positive control.
    Purification
    Affinity Purified
    Immunogen
    The immunogen is a synthetic peptide directed towards the N terminal region of human SPNS2
    Top Product
    Discover our top product SPNS2 Primary Antibody
  • Application Notes
    Optimal working dilutions should be determined experimentally by the investigator.
    Comment

    Antigen size: 549 AA

    Restrictions
    For Research Use only
  • Validation #101829 (Western Blotting)
    'Independent Validation' Badge
    by
    Puissant lab, Institute of Hematology, Inserm U944, CNRS UMR 7212
    No.
    #101829
    Date
    02/01/2018
    Antigen
    SPNS2
    Lot Number
    QC66273-42972
    Method validated
    Western Blotting
    Positive Control
    Baf-3 cells/ MLL-AF9 murine primary cell model
    Negative Control
    Baf-3 or MLL-AF9 murine primary cell model silenced for SPNS2 gene with 2 different shRNA
    Notes

    Passed. ABIN2786494 specifically recognizes SPNS2 in murine Baf-3 cells or MLL-AF9 primary cell model.

    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN2786494
    Secondary Antibody
    mouse anti-rabbit HRP conjugated antibody (GE Healthcare, NA9340V)
    Full Protocol
    • Grow Baf-3 or MLL-AF9 cells in RPMI 1640 1x (Gibco, 21875-059) supplemented with 10% FBS (Sigma-Aldrich, F6178, lot 16G148) and 1x Penicillin/Streptomycin (Gibco, 15140-122), at 37°C and 5% CO2 in 10ml on a flask to 70% confluency.
    • Transduce 3x106 cells with 35µl of concentrated retrovirus, spin-infection (3h at 37°C to 1200 rpm).
    • Grow cells for 3d.
    • Induce and select cells.
    • Grow cells for 5d.
    • Sort and collect cells (doxycycline).
    • Lyse 7x105 cells in 25µl per well in cold SDS Lysis Buffer (Cell Signaling).
    • Determine total protein content of the lysates using Bradford assay (Bio-Rad, 5000006).
    • Denature 20µg of total protein for 7min at 95°C in 6µl Laemmli SDS sample buffer and subsequently separate them on a denaturing NuPAGE 4-12% Bis-Tris Protein Gels (Invitrogen,NP0321BOX ) in an ELECTROPHORESIS CHAMBER for 2h at 150V.
    • Transfer proteins onto PVDF membrane (Millipore) with a Western blotting system for 1h 40min at 300mA.
    • Block the membrane with blocking buffer (2.75% gelatine, 30% BSA, 10mM Tris, NaCl, EDTA, tween 20) for 1h at RT.
    • Incubation with rabbit anti-SPNS2 antibody (antibodies-online, ABIN2786494, lot QC66273-42972) diluted 1:500 in blocking buffer for 16h at 4°C.
    • Wash membrane 3x for 10min with tween TBS buffer.
    • Incubation with secondary mouse anti-rabbit HRP conjugated antibody (GE Healthcare, NA9340V) diluted 1:5000 in blocking buffer for 1h at RT.
    • Wash membrane 3x for 10min with TBST buffer.
    • Reveal protein bands using Pierce ECL Western Blotting Substrate (ThermoFisher Scientific, 32106) on a Li-Cor imaging machine.
    Experimental Notes
    • The SPNS2 antibody ABIN2786494 reveals a protein of the expected molecular weight for SPNS2 in lysates of murine Baf-3 cells or MLL-AF9 primary cell model.

    • ABIN2786494 does show some unspecific binding but the SPNS2 band itself is clear and only visible in the positive but not the negative controls.

  • Format
    Liquid
    Concentration
    Lot specific
    Buffer
    Liquid. Purified antibody supplied in 1x PBS buffer with 0.09 % (w/v) sodium azide and 2 % sucrose.
    Preservative
    Sodium azide
    Precaution of Use
    This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Handling Advice
    Avoid repeated freeze-thaw cycles.
    Storage
    -20 °C
    Storage Comment
    For short term use, store at 2-8°C up to 1 week. For long term storage, store at -20°C in small aliquots to prevent freeze-thaw cycles.
  • Target
    SPNS2 (Spinster Homolog 2 (SPNS2))
    Alternative Name
    SPNS2 (SPNS2 Products)
    Synonyms
    fi20h04 antibody, si:dkey-7b17.4 antibody, toh antibody, wu:fi20h04 antibody, DKFZp459J1933 antibody, spinster homolog 2 (Drosophila) antibody, sphingolipid transporter 2 antibody, spinster homolog 2 antibody, SPNS2 antibody, spns2 antibody, Spns2 antibody
    Background
    SPNS2 is the sphingolipid transporter required for migration of myocardial precursors. SPNS2 transports sphingosine 1-phosphate (S1P), a secreted lipid mediator that plays critical roles in cardiovascular, immunological, and neural development and function. SPNS2 mediates the export of S1P from cells in the extraembryonic yolk syncytial layer (YSL), thereby regulating myocardial precursor migration.
    Alias Symbols: -
    Protein Size: 549
    Molecular Weight
    60 kDa
    Gene ID
    124976
    NCBI Accession
    NM_001124758, NP_001118230
    UniProt
    Q8IVW8
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