Anti-AIM2 Antibody Picoband®

Western blot analysis of AIM2 using anti-AIM2 antibody (PB9683).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human A549 whole cell lysates,
Lane 2: human Daudi whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human HepG2 whole cell lysates,
Lane 5: mouse testis tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-AIM2 antigen affinity purified polyclonal antibody (Catalog # PB9683) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for AIM2 at approximately 45 kDa. The expected band size for AIM2 is at 39,40-45 kDa.

Gene expression and cellular distribution of NLRP1, NLRP3 and Aim2 sensors in the retina. (A) RNAscope analysis of NLRP1 (green dots, yellow arrows) and NLRP3 (white dots, white arrows) transcript abundance in normotensive control (NT control) and experimental (OHT) retinas at 12 h postinjury. (B) Aim2 transcript (red dots) abundance in normotensive control and experimental retinas. Co-staining for the Muller glia marker glutamine synthetase (GlSyn, green) is added to indicate cellular expression of Aim2 in the GCL layer (yellow arrows) and INL (white arrows) in control and 12 h post-OHT retinas. Bar, 25 μm on all panels.
Index in PubMed under a CC BY license. PMID: 30930743

OHT-induced upregulation of Aim2 inflammasome in the retina. (A) NLRP1 antibody labeling (red) in the inner retina; yellow arrows indicate colocalization with RGC neurons (RBPSM + ) in the GCL. (B) Immunohistochemistry for Aim2 protein (red) and Muller glia marker (GlSyn, green). Arrows show colocalization with Muller glia; no colocalization is detected with astrocytes (GFAP, white) in both control and 24 h post-OHT retinas. (C) NLRP3 labeling (green) in the inner retina of naïve and control eyes localized only to blood capillaries (white arrowhead) and astrocytes (yellow arrows), but not to RGCs (RBPMS cells). At 24 h post-injury the labeling in large cells in the GCL and INL (white arrows) are observed. (D) The NLRP1and NLRP3 proteins localized to distinct cell types in the GCL of naïve and NT control retinas: NLRP3 (green) is only expressed in blood vessels (white arrowheads). Twenty four hours after OHT, NLRP3 colocalizes with NLRP1 in RGCs (yellow arrows) and to RBPMS- cells (white arrows). Blue arrowheads denote labeling in RGC axons and dendrites. Bar, 25 μm on all panels.
Index in PubMed under a CC BY license. PMID: 30930743

IHC analysis of AIM2 using anti-AIM2 antibody (PB9683).
AIM2 was detected in a paraffin-embedded section of mouse kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-AIM2 Antibody (PB9683) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

IHC analysis of AIM2 using anti-AIM2 antibody (PB9683).
AIM2 was detected in a paraffin-embedded section of rat intestine tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-AIM2 Antibody (PB9683) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

IHC analysis of AIM2 using anti-AIM2 antibody (PB9683).
AIM2 was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-AIM2 Antibody (PB9683) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.