Sentence examples for min with secondary from inspiring English sources

After three consecutive washes in wash buffer, cells were incubated for 45 min with secondary antibodies (Alexa Fluor Donkey-anti-mouse and Alexa Fluor Donkey-antInvitrogenInvitogethertogether with Hoechst 33342 (Invitrogen).

Membranes were washed three times with 5%% milk TTBS, incubated for 60 min with secondary mouse antibodies conjugated to peroxidase (1 3000; Thermo Scientific, Il, USA), and washed again with TTBS to reveal reactivity with the enhanced chemiluminescence (ECL) reagent (Denville Scientific, NJ, USA).

After blocking with 0.1% BSA in PBS, the samples were incubated in dark conditions for 45 min with secondary antibodies: Alexa 555 donkey anti-mouse (1 500; Invitrogen, Life Technologies, Carlsbad, CA, USA) and Alexa 488 donkey anti-rabbit (1 500, Invitrogen).

After three washes with PBS, the cells were incubated for 30 min with secondary antibodies and DAPI.

Sections were fixed for 3 min in acetone/methanol (1 1) at −20°C, stained for 24 h at 4°C with primary antibody and then incubated 45 min with secondary antibodies.

Binding of primary antibodies was detected by incubating the sections for 30 min with secondary antibodies diluted in PBS with normal rat serum: rabbit anti-mouse HRP (DAKO, Belgium), goat anti-rabbit FITC (SouthernBiotech, USA), and rabbit anti-sheep HRP (DAKO).

The membranes were incubated in primary antibody for 60 min at room temperature, washed with TBS-t (2×5 min), incubated with secondary antibody for 60 min, washed with TBS-t (2×5 min), and finally washed with TBS (1×15 min).

The membrane was probed with the indicated primary antibody, washed three times for 30 min, incubated with secondary antibody conjugated to horseradish peroxidase for 2 h, and washed three times for 30 min.

Talin staining was revealed after 30 min incubation with secondary antibodies conjugated to AlexaFluor 488.

2× washing was followed by 30 min incubation with secondary antibodies conjugated to either Cy3 (1∶500, Jackson Lab) or Alexa-488, -555, -647 (1∶1000, Molecular probes, Invitrogen).

After two washes in buffer A, a 45 min incubation with secondary antibody in buffer A with 5% NGS and 0.1% saponin was performed.