重组兔IgG,单克隆抗体[EPR25A] -同型对照(ab172730)

ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/µL, 2.5 x 10^5 HeLa (Human cervix adenocarcinoma epithelial cell line) cells and 0.5 µg, 1 µg or 2µg of ab172730 [EPR25A]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. H3K4me3 (ab213224) used for comparison.

Additional screenshots of mapped reads can be downloaded here.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

Immunohistochemical analysis of paraffin-embedded human colon tissue using anti-Vimentin RabMAb (ab92547, left panel) (brown) and Rabbit mAb IgG control (ab172730, right panel).

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.

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.

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).

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.

Immunohistochemical analysis of paraffin-embedded mouse lung tissue using anti-Vimentin RabMAb (ab92547, left panel) (brown) and Rabbit mAb IgG control (ab172730, right panel).

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue with unpurified Rabbit IgG ab172730 at 1/10. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue with purified Rabbit IgG ab172730 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.

Overlay histogram showing SH-SY5Y (human neuroblastoma) cells stained with ab179513 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with ab179513 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.

Overlay histogram showing A549 (human lung carcinoma) cells stained with ab185633 (red line). The cells were fixed with 2% paraformaldehyde. The cells were then incubated with ab185633 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

Immunofluorescent staining of HeLa cells using anti-AIF RabMAb (ab32516, left panel) (green) and Rabbit mAb IgG control (ab172730, right panel). DAPI nuclear staining (blue).

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

Immunocytochemistry/immunofluorescence analysis of HeLa cells with purified Rabbit IgG ab172730 at 1/100. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).

Immunocytochemistry/immunofluorescence analysis of HeLa cells with unpurified Rabbit IgG ab172730 at 1/10. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/200) was used as the secondary antibody. Counterstained with DAPI (blue).