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Product Info Summary
| SKU: | A00495-2 |
|---|---|
| Size: | 100 μg/vial |
| Reactive Species: | Human |
| Host: | Rabbit |
| Application: | ELISA, Flow Cytometry, IHC, WB |
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Product info
Product Name
Anti-CARD4/NOD1 Antibody Picoband®
SKU/Catalog Number
A00495-2
Size
100 μg/vial
Form
Lyophilized
Description
Boster Bio Anti-CARD4/NOD1 Antibody Picoband® catalog # A00495-2. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Human. 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
Store 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 freeze-thaw cycles.
Cite This Product
Anti-CARD4/NOD1 Antibody Picoband® (Boster Biological Technology, Pleasanton CA, USA, Catalog # A00495-2)
Host
Rabbit
Contents
Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.
Clonality
Polyclonal
Isotype
Rabbit IgG
Immunogen
E.coli-derived human CARD4/NOD1 recombinant protein (Position: N26-F953).
Cross-reactivity
No cross-reactivity with other proteins.
Reactive Species
A00495-2 is reactive to NOD1 in Human
Observed Molecular Weight
107 kDa
Calculated molecular weight
107.7 kDa
Background of NOD1
Nucleotide-binding oligomerization domain-containing protein 1, also known as CARD4, is a protein receptor that in humans is encoded by the NOD1 gene. NOD1 is a member of NOD-like receptor protein family and is a close relative of NOD2. NOD1 is mapped to 7p14.3. It recognizes bacterial molecules and stimulates an immune reaction. NOD1 protein contains a caspase recruitment domain (CARD). This gene is an intracellular pattern recognition receptor, which is similar in structure to resistant proteins of plants, and mediates innate and acquired immunity by recognizing bacterial molecules containing D-glutamyl-meso-diaminopimelic acid (iE-DAP) moiety. What wore, it has been shown that NOD1 can sense cytosolic microbial products by monitoring the activation state of small Rho GTPases.
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
A00495-2 is guaranteed for ELISA, Flow Cytometry, IHC, 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, 0.25-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human
ELISA, 0.1-0.5μg/ml, -
Positive Control
WB: human THP-1 whole cell, human U20S whole cell, human HELA whole cell
IHC: human liver cancer tissue
FCM: CACO-2 cell
Validation Images & Assay Conditions
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Western blot analysis of CARD4/NOD1 using anti-CARD4/NOD1 antibody (A00495-2).
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 50ug of sample under reducing conditions.
Lane 1: human THP-1 whole cell lysates,
Lane 2: human U20S whole cell lysates,
Lane 3: human HELA whole cell lysates.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-CARD4/NOD1 antigen affinity purified polyclonal antibody (Catalog # A00495-2) 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 CARD4/NOD1 at approximately 107KD. The expected band size for CARD4/NOD1 is at 107KD.
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Flow Cytometry analysis of CACO-2 cells using anti-CARD4/NOD1 antibody (A00495-2).
Overlay histogram showing CACO-2 cells stained with A00495-2 (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with rabbit anti-CARD4/NOD1 Antibody (A00495-2, 1μg/1x106 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x106) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.
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IHC analysis of CARD4/NOD1 using anti-CARD4/NOD1 antibody (A00495-2).
CARD4/NOD1 was detected in paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2μg/ml rabbit anti-CARD4/NOD1 Antibody (A00495-2) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.
Specific Publications For Anti-CARD4/NOD1 Antibody Picoband® (A00495-2)
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