Rabbit anti-Human CARD4/NOD1 Polyclonal Antibody | anti-NOD1 antibody

Anti-CARD4/NOD1 Antibody

Cat. Num. AAA1753333

• Gene Names: NOD1; CARD4; NLRC1; CLR7.1
• Reactivity: Human
• Applications: Western Blot, Immunohistochemistry, Flow Cytometry, Functional Assay, ELISA
• Purity: Immunogen affinity purified.
• Synonyms: CARD4/NOD1; Polyclonal Antibody; Anti-CARD4/NOD1 Antibody; NOD1; CARD4; Nucleotide-binding oligomerization domain-containing protein 1; Caspase recruitment domain-containing protein 4; nucleotide binding oligomerization domain containing 1; anti-NOD1 antibody

• Host: Rabbit
• Reactivity: Human
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Specificity: Rabbit IgG polyclonal antibody for CARD4/NOD1 detection.
• Purity/Purification: Immunogen affinity purified.
• Form/Format: Lyophilized. Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.

• Applicable Applications for anti-NOD1 antibody: Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA)
• Application Notes: WB: 0.25-0.5ug/ml|Human|; IHC-P: 2-5ug/ml|Human|; FC/FACS/FCM: 1-3ug/1x106 cells|Human|; Direct ELISA: 0.1-0.5ug/ml|Human|
• Immunogen: E Coli-derived human CARD4/NOD1 recombinant protein (Position: N26-F953).
• Reconstitution: Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
• Recommended Detection Systems: Recommended Detection Systems
• Preparation and Storage: Store at -20 degree C for one year. After reconstitution, at 4 degree C for one month. It can also be aliquotted and stored frozen at -20 degree C for a longer time. Avoid repeated freezing and thawing.

Flow Cytometry (FC/FACS)

(Figure 2. Flow Cytometry analysis of CACO-2 cells using anti-CARD4/NOD1 antibody (MBS1753333).Overlay histogram showing CACO-2 cells stained with MBS1753333 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CARD4/NOD1 Antibody (MBS1753333, 1μg/1x106 cells) for 30 min at 20 degree C. DyLight®488 conjugated goat anti-rabbit IgG (5-10μg/1x106 cells) was used as secondary antibody for 30 minutes at 20 degree 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. )

Western Blot (WB)

(Figure 1. Western blot analysis of CARD4/NOD1 using anti-CARD4/NOD1 antibody (MBS1753333).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 lysatesLane 2: human U20S whole cell lysatesLane 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 # MBS1753333) at 0. 5 μg/mL overnight at 4 degree 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 # MBS176460) 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. )

Immunohistochemistry (IHC)

(Figure 3. IHC analysis of CARD4/NOD1 using anti-CARD4/NOD1 antibody (MBS1753333).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 (MBS1753333) overnight at 4 degree C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37 degree C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # MBS176451) with DAB as the chromogen. )

Related Product Information for anti-NOD1 antibody

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.

References

1. Chamaillard, M., Hashimoto, M., Horie, Y., Masumoto, J., Su, Q., Saab, L., Ogura, Y., Kawasaki, A., Fukase, K., Kusumoto, S., Valvano, M. A., Foster, S. J., Mak, T. W., Nunez, G., Inohara, N. An essential role for NOD1 in host recognition of bacterial peptidoglycan containing diaminopimelic acid. Nature Immun. 4: 702-707, 2003.
2. da Silva Correia, J., Miranda, Y., Austin-Brown, N., Hsu, J., Mathison, J., Xiang, R., Zhou, H., Li, Q., Han, J., Ulevitch, R. J. Nod1-dependent control of tumor growth. Proc. Nat. Acad. Sci. 103: 1840-1845, 2006.
3. Martinon F, Tschopp J (2006). "NLRs join TLRs as innate sensors of pathogens. ". Trends Immunol. 26 (8): 447-54.

NCBI and Uniprot Product Information

• NCBI GI #: 5174617
• NCBI GeneID: 10392
• NCBI Accession #: NP_006083.1
• NCBI GenBank Nucleotide #: NM_006092.2
• UniProt Accession #: Q9Y239; Q549U4; Q8IWF5
• Molecular Weight: 953
• NCBI Official Full Name: nucleotide-binding oligomerization domain-containing protein 1
• NCBI Official Synonym Full Names: nucleotide-binding oligomerization domain containing 1
• NCBI Official Symbol: NOD1
• NCBI Official Synonym Symbols: CARD4; NLRC1; CLR7.1
• NCBI Protein Information: 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 rep
• UniProt Protein Name: Nucleotide-binding oligomerization domain-containing protein 1
• Protein Family: Nucleotide-binding oligomerization domain-containing protein
• UniProt Gene Name: NOD1
• UniProt Synonym Gene Names: CARD4
• UniProt Entry Name: NOD1_HUMAN