VALIDATION IMAGES
Formalin-fixed and paraffin-embedded rat brain labeled with AIF1 Polyclonal Antibody at 1:200 dilution followed by incubation with secondary antibody and DAB staining
Formalin-fixed and paraffin embedded rat brain tissue labeled with Anti-AIF1 Polyclonal Antibody, unonjugated (bs-1363R) at 1:200 for 40 minutes at 37°C followed by labeling Goat Anti-Rabbit IgG, Cy3 conjugated(bs-0295G-Cy3) 1:200, 40 minutes at 37°C
Paraformaldehyde-fixed, paraffin embedded Rat liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AIF1 (9A3) Polyclonal Antibody, Unconjugated(bs-1363R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.
araformaldehyde-fixed, paraffin embedded rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AIF1 (9A3) Polyclonal Antibody, Unconjugated(bs-1363R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.
BV-microglia stained with AIF1 Polyclonal Antibody (bs-1363R) at 1:100 dilution. AIF1 expression (green) in activated BV-microglia is observed using confocal microscopy. The blue staining represents DAPI. (a) Control; (b) treated with CCL2 (3 ng/mL); (c) CCL2 (3 ng/mL) + l-CDL (3 µM); (d) CCL2 (3 ng/mL) + l-CDL (10 µM); (e) CCL2 (3 ng/mL) + l-CDL (30 µM); (f) CCL2 (3 ng/mL) + RS504393 (10 µM);
Paraformaldehyde-fixed, paraffin embedded Rat Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AIF1 (9A3) Polyclonal Antibody, Unconjugated(bs-1363R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AIF1 (9A3) Polyclonal Antibody, Unconjugated(bs-1363R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat small Intestine;; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AIF1 (9A3) Polyclonal Antibody, Unconjugated(bs-1363R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AIF1 (9A3) Polyclonal Antibody, Unconjugated(bs-1363R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.
U937 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with AIF1 Polyclonal Antibody(bs-1363R-AF488)at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
Paraformaldehyde-fixed, paraffin embedded Human Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AIF1 (9A3) Polyclonal Antibody, Unconjugated(bs-1363R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AIF1 (9A3) Polyclonal Antibody, Unconjugated(bs-1363R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AIF1 (9A3) Polyclonal Antibody, Unconjugated(bs-1363R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining
Paraformaldehyde-fixed, paraffin embedded Mouse Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AIF1 (9A3) Polyclonal Antibody, Unconjugated(bs-1363R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AIF1 (9A3) Polyclonal Antibody, Unconjugated(bs-1363R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining
Paraformaldehyde-fixed, paraffin embedded Mouse Spleen; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AIF1 (9A3) Polyclonal Antibody, Unconjugated(bs-1363R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Small Intestine; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with AIF1 (9A3) Polyclonal Antibody, Unconjugated(bs-1363R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit(Rabbit, SP-0023) and DAB (C-0010) staining.
The THP-1 (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20℃,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.).Primary Antibody (green):Rabbit Anti-AIF1/Iba1 antibody (bs-1363R): 1 μg/10^6 cells; Secondary Antibody (white blue): Goat anti-Rabbit IgG-FITC (bs-40295G-FITC): 1 μg/test. Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
4% Paraformaldehyde-fixed THP-1 (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (AIF1/Iba1) polyclonal Antibody, unconjugated (bs-1363R) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-40295G-FITC) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
25 ug total protein per lane of various lysates (see on figure) probed with AIF1 (9A3) polyclonal antibody, unconjugated (bs-1363R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.