ASC Antibody

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ProSci
Product Code: PSI-2287
Product Group: Primary Antibodies
Supplier: ProSci
CodeSizePrice
PSI-2287-0.02mg0.02mg£150.00
Quantity:
PSI-2287-0.1mg0.1mg£449.00
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Overview

Host Type: Rabbit
Antibody Isotype: IgG
Antibody Clonality: Polyclonal
Regulatory Status: RUO
Applications:
  • Enzyme-Linked Immunosorbent Assay (ELISA)
  • Immunohistochemistry (IHC)
  • Western Blot (WB)

Images

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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.
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<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.
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<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.
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<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.
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<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).
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<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.
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<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.
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<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.
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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.

Documents

Further Information

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
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.
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 References:
  • Masumoto et al. J Biol Chem. 1999;274(48):33835-8.
  • Masumoto et al. Exp Cell Res. 2001;262(2):128-133.
  • Conway et al. Cancer Res. 2000;60(22):6236-42
  • McConnell et al. Cancer Res. 2000;60(22):6243-7. (WD0102)
Buffer:
ASC Antibody is supplied in PBS containing 0.02% sodium azide.
Concentration:
1 mg/mL
Conjugate:
Unconjugated
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.
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.
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.
NCBI Gene ID #:
29108
NCBI Official Name:
PYD and CARD domain containing
NCBI Official Symbol:
PYCARD
NCBI Organism:
Homo sapiens
Physical State:
Liquid
PREDICTED MOLECULAR WEIGHT:
Predicted: 22kD

Observed: 22 kD
Protein Accession #:
BAA87339
Protein GI Number:
10801602
Purification:
ASC Antibody is affinity chromatography purified via peptide column.
Research Area:
Apoptosis
Swissprot #:
Q9ULZ3
User NOte:
Optimal dilutions for each application to be determined by the researcher.
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. 

References

  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

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