Product Info Summary
SKU: | A00362-5 |
---|---|
Size: | 100 µg/vial |
Reactive Species: | Mouse |
Host: | Rabbit |
Application: | ELISA, Flow Cytometry, IF, ICC, WB |
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Product info
Product Name
Anti-ASC/TMS1/Pycard Antibody Picoband™
SKU/Catalog Number
A00362-5
Size
100 µg/vial
Form
Lyophilized
Description
Boster Bio Anti-ASC/TMS1/Pycard Antibody Picoband™ catalog # A00362-5. Tested in ELISA, Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Mouse.
Storage & Handling
At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.
Cite This Product
Anti-ASC/TMS1/Pycard Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00362-5)
Host
Rabbit
Contents
Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived mouse ASC/TMS1/Pycard recombinant protein (Position: K24-S193).
*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00362-5 is reactive to PYCARD in Mouse
Applications
A00362-5 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee
Observed Molecular Weight
22 kDa
Calculated molecular weight
21.627kDa
Background of ASC/TMS1
PYCARD, often referred to as ASC (Apoptosis-associated speck-like protein containing a CARD), is a protein that in humans is encoded by the PYCARD gene. This gene encodes an adaptor protein that is composed of two protein-protein interaction domains: a N-terminal PYRIN-PAAD-DAPIN domain (PYD) and a C-terminal caspase-recruitment domain (CARD). The PYD and CARD domains are members of the six-helix bundle death domain-fold superfamily that mediates assembly of large signaling complexes in the inflammatory and apoptotic signaling pathways via the activation of caspase. In normal cells, this protein is localized to the cytoplasm; however, in cells undergoing apoptosis, it forms ball-like aggregates near the nuclear periphery. Two transcript variants encoding different isoforms have been found for this gene.
Antibody Validation
Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.
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Assay dilution & Images
Reconsitution
Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.
Assay Dilutions Recommendation
The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.
Western blot, 0.25-0.5 µg/ml, Mouse
Immunocytochemistry/Immunofluorescence, 5 µg/ml, Mouse
Flow Cytometry, 1-3 µg/1x1x106 cells, Mouse
Direct ELISA, 0.1-0.5 µg/ml, Mouse
Validation Images & Assay Conditions
Click image to see more details
Figure 1. Western blot analysis of ASC/TMS1/Pycard using anti-ASC/TMS1/Pycard antibody (A00362-5).
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: mouse lung tissue lysates,
Lane 2: mouse spleen tissue lysates,
Lane 3: mouse thymus 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-ASC/TMS1/Pycard antigen affinity purified polyclonal antibody (Catalog # A00362-5) 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 ASC/TMS1/Pycard at approximately 22 kDa. The expected band size for ASC/TMS1/Pycard is at 22 kDa.
Click image to see more details
Figure 2. IF analysis of ASC/TMS1/Pycard using anti-ASC/TMS1/Pycard antibody (A00362-5) and anti-Tubulin Alpha antibody (M03989-3).
ASC/TMS1/Pycard was detected in immunocytochemical section of RM1 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL rabbit anti-ASC/TMS1/Pycard Antibody (A00362-5) and mouse anti-Tubulin Alpha antibody (M03989-3) overnight at 4°C. Cy3 Conjugated Goat Anti-Rabbit IgG (BA1032) and DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) were used as secondary antibody at 1:500 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.
Click image to see more details
Figure 3. Flow Cytometry analysis of RAW264.7 cells using anti-ASC/TMS1/Pycard antibody (A00362-5).
Overlay histogram showing RAW264.7 cells stained with A00362-5 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-ASC/TMS1/Pycard Antibody (A00362-5, 1 μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x106) used under the same conditions. Unlabelled sample (Red line) was also used as a control.
Protein Target Info & Infographic
Gene/Protein Information For PYCARD (Source: Uniprot.org, NCBI)
Gene Name
PYCARD
Full Name
Apoptosis-associated speck-like protein containing a CARD
Weight
21.627kDa
Alternative Names
ASC; ASCMGC10332; CARD5; CARD5TMS; caspase recruitment domain protein 5; Caspase recruitment domain-containing protein 5; hASC; PYCARD; PYD and CARD domain containing; PYD and CARD domain-containing protein; Target of methylation-induced silencing 1; TMS1; TMS-1; TMS1apoptosis-associated speck-like protein containing a CARD PYCARD ASC, CARD5, TMS, TMS-1, TMS1 PYD and CARD domain containing apoptosis-associated speck-like protein containing a CARD|caspase recruitment domain-containing protein 5|target of methylation-induced silencing 1
*If product is indicated to react with multiple species, protein info is based on the gene entry specified above in "Species".For more info on PYCARD, check out the PYCARD Infographic
We have 30,000+ of these available, one for each gene! Check them out.
In this infographic, you will see the following information for PYCARD: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].
Specific Publications For Anti-ASC/TMS1/Pycard Antibody Picoband™ (A00362-5)
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