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TIAM1 anticorps (AA 1-378)

TIAM1 Reactivité: Souris WB Hôte: Mouton Polyclonal unconjugated
N° du produit ABIN1982879
  • Antigène Voir toutes TIAM1 Anticorps
    TIAM1 (T-Cell Lymphoma Invasion and Metastasis 1 (TIAM1))
    Épitope
    • 14
    • 11
    • 5
    • 3
    • 2
    • 2
    • 1
    • 1
    • 1
    AA 1-378
    Reactivité
    • 32
    • 6
    Souris
    Hôte
    • 30
    • 1
    • 1
    Mouton
    Clonalité
    • 31
    • 1
    Polyclonal
    Conjugué
    • 18
    • 4
    • 3
    • 3
    • 2
    • 2
    Cet anticorp TIAM1 est non-conjugé
    Application
    • 25
    • 18
    • 7
    • 3
    • 3
    • 3
    • 3
    • 3
    • 1
    • 1
    Western Blotting (WB)
    Attributs du produit
    Polyclonal Sheep anti Mouse TIAM1
    Immunogène
    AA 1-378
    Isotype
    IgG
    Top Product
    Discover our top product TIAM1 Anticorps primaire
  • Indications d'application
    Optimal dilutions should be determined by each laboratory for each application.
    Restrictions
    For Research Use only
  • Validation #100102 (Western Blotting)
    'Independent Validation' signe
    by
    Instituto de Parasitología y Biomedicina López-Neyra
    No.
    #100102
    Date
    23.12.2016
    Antigène
    TIAM1
    Numéro du lot
    2016090702
    Application validée
    Western Blotting
    Contrôle positif
    primary mouse microglia, MCF-7
    Contrôle négative
    Conclusion
    Passed. ABIN1982879 specifically recognizes TIAM1 in lysates from human MCF-7 cells.
    'Independent Validation' signe
    Validation Images
    Protocole
    Anticorps primaire
    ABIN1982879
    Anticorps secondaire
    anti-goat/sheep HRP-conjugated antibodies (DakoCytomation, P0449, lot 00062104)
    Full Protocol
    • Primary mouse microglia and human MCF-7 cells are grown in DMEM (Invitrogen), supplemented with 10% foetal bovine serum (Gibco), 10% horse serum and penicillin and streptomycin (Gibco), at 37°C and 5% CO2 in a 12-well plate or in RPMI 1640 (Invitrogen), supplemented with 6% inactivated foetal bovine serum (Gibco), GlutaMax (Gibco), penicillin and streptomycin (Gibco), at 37°C and 5% CO2 in a 10cm dish, respectively.
    • Lyse microglia in 25µl per well and MCF-7 cells in 500µl per dish cold lysis buffer (10mM Tris-HCl pH 8.0, 150mM NaCl, 1% Nonidet-P40, 1mM EDTA, 10mM NaF, 1mM Na3VO4, containing protease inhibitors (Sigma)).
    • Determine total protein content of the lysates using Bradford assay (Bio-Rad, 500-0006, lot 111832).
    • Denature 30µg total protein for 5min at 95°C in in 20µl Laemmli SDS sample buffer and subsequently separate them on a denaturing, freshly cast 7.5% SDS-PAGE for 1h at 140V.
    • Transfer proteins onto PVDF membrane (Pall Life Sciences, 75696G, lot TO3225) with a wet Western blotting system for 80min at 100V.
    • Block the membrane with TBST (TBS, 0.1% Tween) containing 5% milk ON at 4°C.
    • Incubation with primary sheep anti-TIAM1 antibody (antibodies-online, ABIN1982879, lot 2016090702) diluted 1:500 in TBST containing 5% milk for 1h at RT.
    • Wash membrane 5x 5min with TBST.
    • Incubation with anti-goat/sheep horseradish peroxidase-conjugated secondary antibodies (DakoCytomation, P0449, lot 00062104) diluted 1:2000 in TBST containing 5% milk for 1h at RT.
    • Wash membrane 5x 5min with TBST and 3x 5min with TBS.
    • Reveal protein bands using self-made ECL Western Blotting Detection Reagent with Curix RP2 Plus medical X-ray films (Agfa, ENKMV, lot 79040074) on an Agfa- Curix60-developer.
    Notes
    The TIAM1 antibody ABIN1982879 reveals a protein of the expected molecular weight of 177kDa in human MCF-7 lysates as verified with a second anti-TIAM1 antibody. However, note that ABIN1982879 did not recognize Tiam1 in primary mouse microglia extracts.
  • Format
    Lyophilized
    Reconstitution
    100 µL of PBS, pH 7.4
    Concentration
    500 µg/mL
    Buffer
    Lyophilized from a solution containing PBS, pH 7.4
    Stock
    -20 °C
  • Antigène
    TIAM1 (T-Cell Lymphoma Invasion and Metastasis 1 (TIAM1))
    Autre désignation
    TIAM1 (TIAM1 Produits)
    Synonymes
    anticorps TIAM1, anticorps tiam1, anticorps AI847750, anticorps D16Ium10, anticorps D16Ium10e, anticorps T-cell lymphoma invasion and metastasis 1, anticorps T-cell lymphoma invasion and metastasis 1a, anticorps T-cell lymphoma invasion and metastasis, anticorps T cell lymphoma invasion and metastasis 1, anticorps TIAM1, anticorps tiam1a, anticorps tiam1, anticorps Tiam1
    UniProt
    Q13009
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