Fluorescein fitc conjugated affinipure goat anti rabbit igg h l

Cells were cultured on glass cover slips, fixed in 4% PFA, and permeabilized with 0.1% Triton X-100 before binding of antibodies recognizing AURKB (Absin, Cat.NO. 131460) and/or human CREST autoimmune serum [14 (link)]. After staining with an appropriate secondary antibody, cells were imaged under an EVOS FL auto microscope. Secondary antibodies were Rhodamine Red^TM-X-conjugated AffiniPure Donkey Anti-Human IgG(H+L) (Jackson, 709-295-149), Fluorescein (FITC)-conjugated Affinipure Goat Anti-Rabbit IgG(H+L) (Jackson, Cat.NO. 111-095-003).

Primary antibodies against AEG-1/MTDH (13860-1-AP), E-cadherin (1702-1), N-cadherin (2447-1), and vimentin (2862-1) were obtained from Epitomics Inc. (Burlingame, CA, USA). Primary antibodies against poly-ADP-ribose polymerase (PARP) (BS70001), PI3K (BS3006), Bax (BS1030), Bcl-2 (BS1031), and C-myc (BS2462) were obtained from Bioworld Technology Inc. (Minneapolis, MN, USA). Primary antibodies against NF-κB-p65 (10745-1-AP), caspase-8 (13423-1-AP), caspase-9 (10380-1-AP), and caspase-3 (19677-1-AP) were obtained from Proteintech (Wuhan, China); Primary antibodies against Lamin A (sc-177452), AKT (sc-8312), p-AKT (sc-7985), ERK (sc-154), and p-ERK (sc-23759) were obtained from Santa Cruz Biotechnology (Santa Cruz Biotechnology Inc., CA). The GAPDH antibody was purchased from Boster (Wuhan, China). The secondary antibodies peroxidase-conjugated AffiniPure Goat Anti-Mouse IgG (H+L) (111-035-003), peroxidase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) (111-035-003), Cy™ 3-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (111-165-003), and Fluorescein (FITC)-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (111-095-003) were obtained from Jackson Immuno Research Laboratories, Inc. (USA). Temozolomide (85622-93-1) was obtained from Meilun Biology Technology (Dalian, China).

Cells grown on coverslips were fixed with acetone, blocked in 10% goat serum in PBS and incubated with antibody against NF-κB P65 (1:50, Cat#8242, Cell Signaling Technology) at 4 °C overnight, followed by 1 hour incubation with Fluorescein (FITC)-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (1:25, Cat#111-095-045, Jackson Immuno Research, PA, USA) at room temperature. Pictures were taken by a laser scanning confocal microscope (LSM 780, Carl Zeiss, Germany)