Table 1.

Primary antibody characteristics

Host/IsotypeAntigen retrievalConcentrationIncubationPositive controlSource
ER-α
    1D5Mouse/IgG1, κHIAR-pH = 6, 15 min7.9 μg/mL1 h, RTMCF-7 cellsDako, Carpinteria, CA
    F10Mouse/IgG2aHIAR-pH = 6, 15 min0.04 μg/mLON, 4°CMCF-7 cellsSanta Cruz, Santa Cruz, CA
    SP1Rabbit/IgGHIAR-pH = 6, 15 min1:1,000*ON, 4°CMCF-7 cellsThermo Fisher, Fremont, CA
    60cRabbit/IgGHIAR-pH = 6, 15 min1:5,000*ON, 4°CMCF-7 cellsMillipore, Billerica, MA
EGFR
    31G7Mouse/IgG1Proteinase K, 5 min RT3 μg/mLON, 4°CEGFR-transfected CHO and A431 cellsZymed/Invitrogen, Carlsbad, CA
    2-18C9MouseProteinase K, 5 min RTPrediluted1 h, RTEGFR-transfected CHO and A431 cellsDako, Carpinteria, CA
    H11Mouse/IgG1, κHIAR-pH = 6, 15 min6.14 μg/mLON, 4°CEGFR-transfected CHO and A431 cellsDako, Carpinteria, CA
    15F8Rabbit/IgGHIAR-pH = 6, 15 min1:50*ON, 4°CEGFR-transfected CHO and A431 cellsCell Signaling, Danvers, MA
    2232Rabbit polyclonalHIAR-pH = 6, 15 min1:200*ON, 4°CEGFR-transfected CHO and A431 cellsCell Signaling, Danvers, MA
HER3
    RTJ2Mouse/IgG1HIAR-pH = 6, 15 min2 μg/mLON, 4°CHER3-transfected BaF3 cellsSanta Cruz, Santa Cruz, CA
    RB_9211Rabbit polyclonalHIAR-pH = 6, 15 min0.2 μg/mLON, 4°CHER3-transfected BaF3 cellsThermo Scientific, Fremont, CA
    C_17Rabbit polyclonalHIAR-pH = 9, 15 min0.2 μg/mLON, 4°CHER3-transfected BaF3 cellsSanta Cruz, Santa Cruz, CA
    M7297Mouse/IgG2a, κHIAR-pH = 9, 15 min13 μg/mLON, 4°CHER3-transfected BaF3 cellsDako, Carpinteria, CA
    SGP1Mouse/IgG1, κHIAR-pH = 6, 15 min4 μg/mLON, 4°CHER3-transfected BaF3 cellsThermo Scientific, Fremont, CA
    RTJ1Mouse/IgMHIAR-pH = 6, 15 min0.5 μg/mLON, 4°CHER3-transfected BaF3 cellsNovocastra, Newcastle upon Tyne, UK

Abbreviations: HIAR, heat-induced antigen retrieval; RT, room temperature; ON, overnight.

  • *The antibody concentration was not provided by the manufacturer.