Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Overlay histogram showing A549 (human lung carcinoma) cells stained with ab133557 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with ab133557 at 1/60 dilution. The secondary antibody used was goat anti-rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of

5,000 events were collected using a 20mW Argon ion laser (488nm) and 530/30 bandpass filter.

Conjugated versions are available for this clone: Alexa Fluor® 488 (ab199091), Alexa Fluor® 647 (ab199093), R-PE (ab209478), APC (ab232814).

Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

ab124962 (purified) at 1/20 immunoprecipitating IL-1RA in NIH/3T3 whole cell lysate.

Lane 1 (input): NIH/3T3 whole cell lysate (10μg)

Lane 2 (+): ab124962 + NIH/3T3 whole cell lysate.

Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab124962 in NIH/3T3 whole cell lysate.

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Blocking buffer and concentration: 5% NFDM/TBST.

Diluting buffer and concentration: 5% NFDM /TBST.

All lanes:

Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Overlay histogram showing K562 (human chronic myelogenous leukemia) cells stained with ab196018 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with ab196018 at 1/150 dilution. The secondary antibody used was goat anti-rabbit IgG (FITC) at 1/150 dilution. Isotype control antibody (black line) was rabbit IgG (monoclonal) (ab172730). Unlabelled control (blue line) was cells without incubation with primary antibody and secondary antibody. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 530/30 bandpass filter.

Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

LINE-1 ORF1p was immunoprecipitated from 0.35 mg F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate with ab216324 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216324 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1: F9 whole cell lysate 10 μg (Input).

Lane 2: ab216324 IP in F9 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216324 in F9 whole cell lysate.

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: 30 seconds.

All lanes:

Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Flow Cytometry - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Flow cytometric analysis of HEK-293T (Human embryonic kidney epithelial cell, Left panel) / Caco-2 (Human colorectal adenocarcinoma epithelial cell, Right panel) using ab278053 at 1/500 dilution (0.1µg) (red). The isotype control used was the Rabbit monoclonal IgG (ab172730), black line and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Secondary antibody used was the Goat anti-rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution.

Negative control: HEK-293T. (PMID 20167130)

Gated on viable cells.

ChIC/CUT&RUN sequencing - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

CUT&RUN was performed using the ChIC/CUT&RUN pAG-MNAse ab285373 105 HeLa cells and 0.5 μg, 1 μg and 2μg of ab172730 [EPR25A]. H3K4me3 (ab213224) and CTCF (ab188408) are used for comparison. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads

Additional screenshots of mapped reads can be downloaded here.