Sentences with phrase «incubated with antibodies»

Precleared nuclear lysates were incubated with antibodies against Nrf2 or Bach1.

The tissue was paraformaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed before the tissue was blocked and incubated with the antibody for 1 hour.

The cells were then incubated with the antibody (ab7291, 1µg / ml) and (ab16048, 1µg / ml) overnight at +4 °C.

The cells were then incubated with the antibody ab16056 at 1µg / ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4 °C.

Not exact matches

The slides were incubated with the primary antibody overnight at 4 °C, rinsed three times with PBS, incubated for 10 min in protein blocking solution, and incubated with the appropriate secondary reagent.

Membranes were incubated overnight at 4 °C with antibodies specific for phospho - Smad1 / 5/8 and Smad1 (Cell Signaling Technology), V5 (Invitrogen), or β - actin (Santa Cruz Biotechnology Inc.) in PBS containing 5 % nonfat milk and 0.5 % BSA.

The SVCs were incubated with a fluorescently labeled anti-F4 / 80 antibody and were sorted by FACS into F4 / 80 - expressing (F4 / 80 +) and - nonexpressing (F4 / 80 ---RRB- populations.

Cells treated with hamster anti-αβTCR or hamster murine γδTCR were washed three times in HBSS and incubated in PE - labeled Armenian hamster antihamster IgG (PharMingen) secondary antibody for 20 min at 4 °C, washed three additional times in PBS, fixed in 1 % paraformaldehyde, and assessed for fluorescence in a FACScan flow cytometer (Becton Dickinson).

The membrane was then incubated with a 1:500 dilution of mouse monoclonal anti-HIF-1α antibody (Novus Biologicals, Littleton, CO) in 5.0 % nonfat dry milk in 0.1 % Tween - 20 in TBS at 4 °C overnight.

The sample was incubated with the primary antibody (1/100 in PBS) for 15 minutes at room temperature.

The sample was incubated with the primary antibody (1/100 in 1x HBSS + 0.02 % Triton X-100 + 1.5 % FBS) for 3 hours at 25 °C.

Following washings, slides were incubated with the respective secondary antibody and Hoechst 1:5,000 at RT for 45 min and mounted with Fluoromount medium (eBioscience).

arabidopsis, rice, wheat, corn whole plant tissue were subjected to SDS PAGE followed by western blot with 60004 -1-Ig (GAPDH Antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours

zebrafish tissue were subjected to SDS PAGE followed by western blot with 60004 -1-Ig (GAPDH antibody) at dilution of 1:6000 incubated at room temperature for 1.5 hours

mouse colon tissue were subjected to SDS PAGE followed by western blot with 11176 -1-AP (HNRNPA1 Antibody) at dilution of 1:600 incubated at room temperature for 1.5 hours

Cells were washed, incubated with the corresponding secondary antibody and Hoechst (Sigma, # 33258, 1:1,000) for 30 min at RT in the dark and coverslips were mounted with Fluoromount G (Dako).

Rat brain tissue lysate was subjected to SDS PAGE followed by western blot with 23449 -1-AP (UBQLN2 antibody) at a dilution of 1:800 incubated at room temperature for 1.5 hours.

In Figure 4, Panel B, cells were incubated simultaneously with anti ‑ SSEA ‑ 4 and anti - TRA ‑ 1 ‑ 60 antibodies, since they were labeled with different fluorophores (PE and FITC, respectively).

After three rinses in TBS, 5 min each, samples were incubated with a secondary antibody.

mouse testis tissue were subjected to SDS PAGE followed by western blot with 10837 -1-AP (OPTN antibody) at dilution of 1:800 incubated at room temperature for 1.5 hours

soybean whole plant tissue were subjected to SDS PAGE followed by western blot with 60004 -1-Ig (GAPDH Antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours

Cell lysates were incubated with protein G sepharose beads (GE Healthcare) and 3 μg Tie2 antibody (Millipore, clone Ab33, # 05 - 584) overnight at 4 °C on a rotator.

After 30 - min incubation with the secondary antibody, tissue sections were incubated with streptavidin conjugated with peroxidase, diluted 1:20 in TBS (Zymed Laboratories, Inc.) for 30 min at room temperature.

Samples were incubated with primary antibody (1/100 in PBST with BSA and goat serum) for 4 °C at 12 hours.

Raji cells were subjected to SDS PAGE followed by western blot with 60004 -1-Ig (GAPDH antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours

A549 cells were subjected to SDS PAGE followed by western blot with 60008 -1-Ig (ACTB antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours

HeLa cells were subjected to SDS PAGE followed by western blot with 11176 -1-AP (HNRNPA1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours

HEK - 293 cells were subjected to SDS PAGE followed by western blot with IFT88 antibody (13967 -1-AP) at a dilution of 1:1000 incubated at room temperature for 1.5 hours.

After washing with PBS and blocking for 30 minutes (IHC / ICC Blocking Buffer - Low Protein; eBioscience), cells were incubated with anti - Oct ‑ 4 antibody (diluted 1:150; eBiosicence) or an IgG2a K isotype control (diluted 1:150; eBiosicence) for one hour.

Confirmation of CD44 positive population post treatment was performed by incubating cell suspension with CD44 primary antibody (Abcam)(1:500 dilution) followed by FITC / TRITC tagged secondary antibody (Abcam)(1:1000 dilution).

MCF7 cells were subjected to SDS PAGE followed by western blot with 60008 -1-Ig (ACTB antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours

mouse brain tissue were subjected to SDS PAGE followed by western blot with 10837 -1-AP (OPTN antibody) at dilution of 1:800 incubated at room temperature for 1.5 hours

arabidopsis whole plant tissue were subjected to SDS PAGE followed by western blot with 60008 -1-Ig (beta actin Antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours

The sections were washed in PBS and incubated for 1 h with a mouse anti-peroxidase monoclonal antibody [37](1:30) pre-incubated with horseradish peroxidase (5 μg / ml) in PBS (MAP kit, Medimabs, Canada).

One pair of slides was incubated with the primary antibody (anti PD - L1 monoclonal mouse antibody, clone 5H1, generated by Dr. Lieping Chen, Yale University, New Haven, CT) diluted at 1:75 in ACE block at 4 °C overnight.

Samples were incubated with primary antibody (1/300 in blocking buffer) for 12 hours at 4 °C.

Total cell lysates of HuH - 7 cells were incubated with GST or GST - Sp1 fusion protein — prebound glutathione - Sepharose 4B beads and the bound fractions were subjected to immunoblotting with antibody against DDX3 (top).

Incubate the membrane with a suitable HRP - conjugated secondary antibody (recognizing the host species of the primary antibody), diluted at 1:5000 — 1:50000 in blocking solution.

rat brain tissue were subjected to SDS PAGE followed by western blot with 21672 -1-AP (SIP1 Antibody) at dilution of 1:1000 incubated at room temperature for 1.5 hours

To examine the evolution of the AD - like amyloid pathology we incubated sections with the monoclonal antibody McSA1 [36](MediMabs, Montreal, Canada) at 1:4000 in PBS - T with 5 % NGS overnight at 4 °C.

The sections were incubated with primary antibody of Cyclin D1 (Genemed Biotechnologies company, South San Francisco, USA) at 4ºC overnight.

Sections were incubated with either (A) no primary antibody or (B) anti-DDX4 (Abcam ab27591) for 1 h at RT..

Sections were incubated with primary antibody in 0.3 % Triton X-100 in KPBS plus 2 % filtered serum or BSA overnight at 4 °C, and with primary antibodies (1:1,000) in 0.3 % Triton X-100 for 1 hr at room temperature.

The following day, the sections were washed in PBS - T and incubated with a goat anti-mouse secondary antibody (MP Biochemicals, Canada) 1:100 in PBS with 5 % NGS for 1 h.

HepG2 cells were subjected to SDS PAGE followed by western blot with 21672 -1-AP (SIP1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours

MCF7 cells were subjected to SDS PAGE followed by western blot with 10379 -1-AP (SNRPD3 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours

Briefly, 1 mL cell suspension (1 × 106 cells) was incubated for few minutes in rat specific Fc block solution (BD Biosciences, San Jose, CA, USA) followed by incubation with fluorescence - tagged primary antibodies - CD11b - FITC (BD Biosciences), TNF - α - PE (BD Biosciences) and NF - κB - FITC (in 1:100 dilution) at 4 °C for 30 min in dark.

HeLa cells were subjected to SDS PAGE followed by western blot with 21672 -1-AP (SIP1 antibody) at dilution of 1:500 incubated at room temperature for 1.5 hours

For double labeling, the cells were incubated with primary antibodies against TuJ1 and L1 or GFAP, then biotinylated anti-mouse or rabbit (rat) IgG (Vector, Burlingame, CA, USA), and finally with avidin - rhodamine (Vector) and corresponding FITC - conjugated secondary antibodies (from different host species, Chemicon and Molecular Probes, Invitrogen).

Incubate sections with primary antibody diluted in 1x TBST for 1 hour, or overnight at 4 °C; the optimal antibody dilution ratio should be pre-determined by experimentation.