PYCARD Antibody

• Catalog Number: orb1239217
• Category: Antibodies
• Description: PYCARD Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, ICC, IHC-P, WB
• Predicted Reactivity: Mouse, Rat
• Reactivity: Human
• Isotype: IgG
• Immunogen: Anti-ASC antibody (orb1239217) was raised against a peptide corresponding to 14 amino acids near the carboxy terminus of human ASC. The immunogen is located within the last 50 amino acids of ASC.
• Concentration: 1 mg/mL
• Dilution range: WB: 1-2 μg/mL; ICC: 5 μg/mL; IHC-P: 2.5 μg/mL.Antibody validated: Western Blot in human samples; Immunocytochemistry in human samples; Immunofluorescence in human samples. All other applications and species not yet tested.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: Predicted: 22kDObserved: 22 kD
• Target: PYCARD
• UniProt ID: Q9ULZ3
• NCBI: BAA87339
• Storage: ASC antibody can be stored at 4°C for three months and -20°C, stable for up to one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.
• Buffer/Preservatives: ASC Antibody is supplied in PBS containing 0.02% sodium azide.
• Alternative names: ASC Antibody: ASC, TMS, TMS1, CARD5, TMS-1, ASC, A
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Western Blot Validation in Human HL60 Cells. Loading: 15 µg of lysates per lane. Antibodies: ASC orb1239217, (1 µg/mL) in the absence (A) or presence (B) of blocking peptide, 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines. Loading: 15 µg of lysates per lane. Antibodies: ASC orb1239217, (2 µg/mL), ASC, (2 µg/mL), beta-actin (1 µg/mL) and GAPDH (0.02 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit or goat anti-mouse (for ASC 39001) IgG HRP conjugate at 1:10000 dilution.

Western Blot Validation in Human THP-1 Cells. Loading: 15 µg of lysate per lane. Antibodies: ASC orb1239217, (2 µg/mL), 1h incubation at RT in 5% NFDM/TBST. Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.

Immunohistochemistry Validation of ASC in Human Spleen Tissue. Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-ASC antibody (orb1239217) at 2.5 µg/ml. Tissue was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

Immunofluorescence Validation of ASC in Human Spleen Tissue. Immunofluorescent analysis of 4% paraformaldehyde-fixed Human Spleen Tissue labeling ASC with orb1239217 at 20 µg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).

Immunocytochemistry Validation of ASC in HL60 Cells. Immunocytochemical analysis of HL60 cells using anti-ASC antibody (orb1239217) at 5 µg/ml. Cells was fixed with formaldehyde and blocked with 10% serum for 1 h at RT; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody overnight at 4°C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin.

KD Validation of ASC in THP-1 Cells (Dowds et al., 2004). Immunofluorescence analysis with anti-ASC antibodies (orb1239217) was performed for BIM in 293 cells transfected with GFP siRNA or ASC siRNA. ASC expression was disrupted after ASC siRNA knockdown.

Overexpression Validation of ASC in HEK293T Cells (Dowds et al., 2004). Western blot analysis with anti-ASC antibodies (orb1239217) was performed for ASC in HEK293T cells transfected with pcDNA3-ASC.