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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.
IAV
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Most Cited
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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 0.02% sodium azide.
Concentration:
1 mg/mL
Storage Conditions:
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.
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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