Recombinant Mouse IgG1 protein [PSH04-91] - Isotype Control

PCNA was immunoprecipitated from 0.2 mg HeLa cell lysate with HA601172 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA601172 at 1/10,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: HeLa cell lysate (input)
Lane 2: HA601172 IP in HeLa cell lysate
Lane 3: Recombinant Mouse IgG1 (HA601336) instead of HA601172 in HeLa cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 1 minute 59 seconds; ECL: K1801

PCNA was immunoprecipitated from 0.2 mg NIH/3T3 cell lysate with HA601172 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA601172 at 1/10,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: NIH/3T3 cell lysate (input)
Lane 2: HA601172 IP in NIH/3T3 cell lysate
Lane 3: Recombinant Mouse IgG1 (HA601336) instead of HA601172 in NIH/3T3 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 5 seconds; ECL: K1802

PCNA was immunoprecipitated from 0.2 mg PC-12 cell lysate with HA601172 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA601172 at 1/10,000 dilution. Anti-Mouse IgG for IP Nano-secondary antibody (NBI02H) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: PC-12 cell lysate (input)
Lane 2: HA601172 IP in PC-12 cell lysate
Lane 3: Recombinant Mouse IgG1 (HA601336) instead of HA601172 in PC-12 cell lysate

Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 43 seconds; ECL: K1801

Flow cytometric analysis of HeLa cells labeling eIF-6.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA601293, 1μg/mL) (green) compared with Recombinant Mouse IgG1 (HA601336, 1μg/mL) (red). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Flow cytometric analysis of C6 cells labeling eIF-6.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA601293, 1μg/mL) (green) compared with Recombinant Mouse IgG1 (HA601336, 1μg/mL) (red). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

☑ Cell treatment (CT)

Chromatin immunoprecipitations were performed with cross-linked chromatin from HeLa cells treated with 500ng/mL TSA for 4 hours with Histone H3 (acetyl K27) (HA600047) or Recombinant Mouse IgG1 (HA601336) according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.