Anti-ADAR1 antibody [EPR7033] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-ADAR1 antibody [EPR7033] - BSA and Azide free (AB240029)

Intracellular flow cytometric analysis of permeabilized Ramos cells, staining ADAR1 (red) with ab126745. 1x10⁶ cells were collected and washed with blocking buffer. Cells were fixed with 2% paraformaldehyde, permeabilized with 1X FACS permeabilizing solution and blocked with blocking buffer for 30 minutes at room temperature. Cells were incubated with primary antibody (1/10) for 30 minutes at room temperature before a Fluorescently-conjugated secondary antibody or 30 min at room temperature. A rabbit IgG was used as a negative control (green). This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126745).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADAR1 antibody [EPR7033] - BSA and Azide free (AB240029)

ab126745, at 1/50 dilution, staining ADAR1 in paraffin-embedded Human brain tissue by Immunohistochemistry.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126745).

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-ADAR1 antibody [EPR7033] - BSA and Azide free (AB240029)

This data was developed using the same antibody clone in a different buffer formulation (ab126745).

Lanes 1-3 : Merged signal (red and green). Green - ab126745 observed at 130 kDa. Red - loading control ab8245 observed at 36 kDa.

ab126745 Anti-ADAR1 antibody [EPR7033] was shown to specifically react with ADAR1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266846 (knockout cell lysate ab257131) was used. Wild-type and ADAR1 knockout samples were subjected to SDS-PAGE. ab126745 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ADAR1 antibody [EPR7033] (<a href='/en-us/products/primary-antibodies/adar1-antibody-epr7033-ab126745'>ab126745</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

ADAR knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human ADAR (ADAR1) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-adar-adar1-knockout-hek-293t-cell-line-ab266846'>ab266846</a>)

Lane 3:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 136 kDa,19 kDa,29 kDa,35 kDa,36 kDa

Observed band size: 130 kDa,20 kDa,29 kDa,35 kDa,36 kDa

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