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<strong>Figure 1 Western Blot Validation in Human HL60 Cells</strong><br>Loading: 15 μg of lysates per lane.Antibodies: ASC 2287, (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.
<strong>Figure 2 Independent Antibody Validation (IAV) via Protein Expression Profile in Cell Lines</strong><br>Loading: 15 μg of lysates per lane.Antibodies: ASC 2287, (2 μg/mL), ASC 39-001, (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.
<strong>Figure 3 Western Blot Validation in Human THP-1 Cells</strong><br>Loading: 15 μg of lysate per lane.Antibodies: ASC 2287, (2 μg/mL), 1h incubation at RT in 5% NFDM/TBST.Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 dilution.
<strong>Figure 4 Immunohistochemistry Validation of ASC in Human Spleen Tissue </strong><br> Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-ASC antibody (2287) 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.
<strong>Figure 5 Immunofluorescence Validation of ASC in Human Spleen Tissue</strong><br>Immunofluorescent analysis of 4% paraformaldehyde-fixed Human Spleen Tissue labeling ASC with 2287 at 20 μg/mL, followed by goat anti-rabbit IgG secondary antibody at 1/500 dilution (red).
<strong>Figure 6 Immunocytochemistry Validation of ASC in HL60 Cells</strong><br>Immunocytochemical analysis of HL60 cells using anti-ASC antibody (2287) 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.
<strong>Figure 7 KD Validation of ASC in THP-1 Cells (Dowds et al., 2004) </strong><br>Immunofluorescence analysis with anti-ASC antibodies (2287) was performed for BIM in 293 cells transfected with GFP siRNA or ASC siRNA. ASC expression was disrupted after ASC siRNA knockdown.
<strong>Figure 8 Overexpression Validation of ASC in HEK293T Cells (Dowds et al., 2004) </strong><br>Western blot analysis with anti-ASC antibodies (2287) was performed for ASC in HEK293T cells transfected with pcDNA3-ASC.
Figure 1 Western Blot Validation in Human HL60 Cells
Loading: 15 μg of lysates per lane.Antibodies: ASC 2287, (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.
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ASC Antibody

CATALOG NUMBER: 2287

Clonality:
Polyclonal
Tested Applications:
ELISA, ICC, IHC-P, WB
Host Species:
Rabbit
Species Reactivity:
Human
Conjugate:
Unconjugated
Specifications
Host Species:
Rabbit
Species Reactivity:
Human
Homology:
Predicted species reactivity based on immunogen sequence: Mouse (57%), Rat (57%)
Immunogen:
Anti-ASC antibody (2287) 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.
Conjugate:
Unconjugated
Tested Applications:
ELISA, ICC, IHC-P, WB
Application Note:
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.
Positive Control 1:
Cat. No. 1209 - HL60 Cell Lysate
Positive Control 2:
Cat. No. 17-009 - HL-60 Cell Slide
Positive Control 3:
Cat. No. 10-901 - Human Spleen Tissue Slide
Positive Control 4:
Cat. No. 1208 – THP-1 Cell Lysate
Predicted Molecular Weight:
Predicted: 22kD
Observed: 22 kD
Validation:

Independent Antibody Validation in Cell lines (Figure 2) shows similar ASC expression profile in human and mouse cell lines detected by two independent anti-ASC antibodies that recognize different epitopes, 2287 against C-terminus domain and 39-001 against full length human recombinant protein.  ASC proteins are detected in some of tested cell lines at different expression levels by the two independent antibodies. 

Isoforms:
Human ASC has 3 isoforms, including isoform 1 (195aa, 22kD), isoform 2 (176aa, 20kD), and isoform 3 (135aa, 15kD). Mouse ASC has one isoform (193aa, 22kD) and rat ASC also has one isoform (193aa, 22kD). 2287 can detect all human isoforms.
Purification:
ASC Antibody is affinity chromatography purified via peptide column.
Clonality:
Polyclonal
Isotype:
IgG
Physical State:
Liquid
Buffer:
ASC Antibody is supplied in PBS containing ≤0.09% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
ASC antibody can be stored at 4°C up to one year. Antibodies should not be exposed to prolonged high temperatures.
Ncbi Official Symbol:
PYCARD
Additional Names:
ASC Antibody: ASC, TMS, TMS1, CARD5, TMS-1, ASC, Apoptosis-associated speck-like protein containing a CARD, Caspase recruitment domain-containing protein 5, hASC
Protein Accession Number:
BAA87339
Protein Gi Number:
10801602
Ncbi Gene Id Number:
29108
User Note:
Optimal dilutions for each application to be determined by the researcher.
Background:
ASC Antibody: Apoptosis is regulated by death domain (DD) and/or caspase recruitment domain (CARD) containing molecules and a caspase family of proteases. CARD containing cell death regulators include RAIDD, RICK, BCL10, Apaf-1, ARC, caspase-9, and caspase-2. A novel CARD domain containing protein was recently identified in human and mouse and designated ASC and TMS1. Ectopic expression of ASC/TMS1 induced apoptosis through activation of caspase-9 and inhibited the survival of human breast cancer cells (3, 4). Overexpression of ASC/TMS1 induced DNA fragmentation. ASC/TMS1 is expressed in a variety of human and mouse tissues.
Background Reference 1:
Masumoto et al. J Biol Chem. 1999;274(48):33835-8.
Background Reference 2:
Masumoto et al. Exp Cell Res. 2001;262(2):128-133.
Background Reference 3:
Conway et al. Cancer Res. 2000;60(22):6236-42
Background Reference 4:
McConnell et al. Cancer Res. 2000;60(22):6243-7. (WD0102)
Citation 1:
Dowds et al. Cryopyrin-induced interleukin 1beta secretion in monocytic cells: enhanced activity of disease-associated mutants and requirement for ASC. J Biol Chem. 2004;279(21):21924-8. PMID: 15020601

FOR RESEARCH USE ONLY

For additional information, visit ProSci’s Terms & Conditions Page.

Disclaimer:
Optimal dilutions/concentrations should be determined by the end user. The information provided is a guideline for product use. This product is for research use only.

CATALOG NUMBER:

2287

List Size:
0.02 mg, 0.1 mg

List Price:

Price range: $99.00 through $445.00

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