Anti-CARD4/NOD1 Antibody Picoband™

Product Name

Anti-CARD4/NOD1 Antibody Picoband™

View all NOD1 Antibodies

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.

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).

*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

A00495-2 is reactive to NOD1 in Human

Applications

A00495-2 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

107 kDa

Calculated molecular weight

107.691kDa

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.

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Reconsitution

Add 0.2ml of distilled water will yield a concentration of 500ug/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, 0.25-0.5μg/ml, Human
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Human
Flow Cytometry, 1-3μg/1x10⁶ cells, Human
Direct ELISA, 0.1-0.5μg/ml, Human

Validation Images & Assay Conditions

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Figure 1. 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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Figure 2. 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 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 (Red line) was also used as a control.

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Figure 3. 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.

Gene/Protein Information For NOD1 (Source: Uniprot.org, NCBI)

Alternative Names

CARD4; CARD4NLR family, CARD domain containing 1; caspase recruitment domain family, member 4; Caspase recruitment domain-containing protein 4; CLR7.1; NLRC1; NOD1; nucleotide-binding oligomerization domain containing 1; nucleotide-binding oligomerization domain, leucine rich repeat and CARD domaincontaining 1; nucleotide-binding oligomerization domain-containing protein 1 NOD1 CARD4, CLR7.1, NLRC1 nucleotide binding oligomerization domain containing 1 nucleotide-binding oligomerization domain-containing protein 1|NLR family, CARD domain containing 1|caspase recruitment domain family, member 4|caspase recruitment domain-containing protein 4|nucleotide-binding oligomerization domain, leucine rich repeat and CARD domain containing 1

For more info on NOD1, check out the NOD1 Infographic

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In this infographic, you will see the following information for NOD1: 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].

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