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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. The brand Picoband indicates this is a premium antibody that guarantees superior quality, high affinity, and strong signals with minimal background in Western blot applications. Only our best-performing antibodies are designated as Picoband, ensuring unmatched performance.
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).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00362-5 is reactive to Pycard in Mouse
Observed Molecular Weight
22 kDa
Calculated molecular weight
21.5 kDa
Background of Pycard
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.
Application & Images
Applications
A00362-5 is guaranteed for ELISA, Flow Cytometry, IF, ICC, WB Boster Guarantee
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 (Fixed), 1-3 μg/1x1x106 cells, Mouse
ELISA, 0.1-0.5 μg/ml, -
Positive Control
WB: mouse lung tissue, mouse spleen tissue, mouse thymus tissue
ICC/IF: RM1 cell
FCM: RAW2647 cell
Validation Images & Assay Conditions
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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.
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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.
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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.
Specific Publications For Anti-ASC/TMS1/Pycard Antibody Picoband® (A00362-5)
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