RPE cells were treated with 1 μM A-1155463, 1 μM 4NQO or both compounds. Control cells were treated with 0.1% DMSO. After 1 and 2 h cells were fixed using 3.3% formaldehyde in 10% FBS and PBS, pH 7.4. Cells were permeabilized using 10% FBS and 0.1% Triton X-100 in PBS. Bad, Bax, Bcl-xL, p53 and Tom40 were detected using rabbit anti-Bcl-xL (1:250; clone 54H6; Cell Signalling Technology), mouse anti-Bax (1:250; 2D2, sc-20067), rabbit anti-Bax (1:250; N-20, sc-493), rabbit anti-BAD (1:250; clone D24A9; Santa Cruz Biotechnology), mouse anti-p53 (1:250; Santa Cruz Biotechnology) and anti-Tom40 (1:250; Santa Cruz Biotechnology, sc-11414) antibodies were used. Secondary goat anti-mouse IgG (H + L) Alexa Fluor® 488 and 568 (Thermo Scientific), goat anti-rabbit IgG (H + L) Alexa Fluor® 488 and 568 (Thermo Scientific) antibodies were used. ProLong™ Gold Antifade Mountant (Invitrogen, Waltham, MA, USA) with DAPI was used for mounting of cells. Cells were imaged using Zeiss LSM710 confocal microscope (Zeiss, Oberkochen, Germany).
Goat anti rabbit igg h l alexa fluor 488 and 568
Manufactured by Thermo Fisher Scientific
Goat anti-rabbit IgG (H + L) Alexa Fluor® 488 and 568 are secondary antibodies labeled with the Alexa Fluor® 488 and 568 fluorophores, respectively. These antibodies are designed to detect and bind to rabbit primary antibodies, recognizing both the heavy (H) and light (L) chains.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 710 confocal microscope, by Zeiss (4 mentions)
Anti tom40, by Santa Cruz Biotechnology (2 mentions)
Clone d24a9, by Santa Cruz Biotechnology (2 mentions)
Goat anti mouse igg h l alexa fluor 488 and 568, by Thermo Fisher Scientific (2 mentions)
Prolong gold antifade mountant, by Thermo Fisher Scientific (2 mentions)
Mouse anti p53, by Santa Cruz Biotechnology (2 mentions)
Rabbit anti bcl xl, by Cell Signaling Technology (1 mentions)
Mouse anti bax, by Santa Cruz Biotechnology (1 mentions)
2 protocols using goat anti rabbit igg h l alexa fluor 488 and 568
1
RPE Cell Apoptosis Evaluation
2
Apoptosis Regulators in RPE Cells
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RPE cells were treated with 1 μM A-1155463, 1 μM 4NQO or both compounds. Control cells were treated with 0,1% DMSO. After 1 and 2 h cells were fixed using 3.3% formaldehyde in 10% FBS and PBS, pH 7.4. Cells were permeabilized using 10% FBS and 0.1% Triton X-100 in PBS. Bad, Bax (2 different antibodies), Bcl-xL, p53 and Tom40 were detected using rabbit anti-Bcl-xL (1 : 250; clone 54H6; Cell Signalling Technology), mouse anti-Bax (1:250; 2D2, sc-20067), rabbit anti-Bax (1:250; N-20, sc-493), rabbit anti-BAD (1:250; clone D24A9; Santa Cruz Biotechnology), mouse anti-p53
(1 : 250; Santa Cruz Biotechnology) and anti-Tom40 (1:250; Santa Cruz Biotechnology, sc-11414) antibodies were used. Secondary goat anti-mouse IgG (H+L) Alexa Fluor ® 488 and 568 (Thermo Scientific), goat anti-rabbit IgG (H+L) Alexa Fluor ® 488 and 568 (Thermo Scientific) antibodies were used. ProLong™ Gold Antifade Mountant (Invitrogen) with DAPI was used for mounting of cells. Cells were imaged using Zeiss LSM710 confocal microscope.
(1 : 250; Santa Cruz Biotechnology) and anti-Tom40 (1:250; Santa Cruz Biotechnology, sc-11414) antibodies were used. Secondary goat anti-mouse IgG (H+L) Alexa Fluor ® 488 and 568 (Thermo Scientific), goat anti-rabbit IgG (H+L) Alexa Fluor ® 488 and 568 (Thermo Scientific) antibodies were used. ProLong™ Gold Antifade Mountant (Invitrogen) with DAPI was used for mounting of cells. Cells were imaged using Zeiss LSM710 confocal microscope.
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