Western Blot (WB)
(Figure 1. Western blot analysis of CARD4 using anti-CARD4 antibody (MBS177898).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: A549 Whole Cell Lysate,Lane 2: Rat Cardiac Muscle Tissue Lysate.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 antigen affinity purified polyclonal antibody at 0.5ug/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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for CARD4 at approximately 107KD. The expected band size for CARD4 is at 107KD. )
anti-Human, Rat CARD4 Polyclonal Antibody | anti-CARD4 antibody
Anti-CARD4 Antibody
Immunohistochemistry (IHC) Paraffin Concentration: 0.5-1ug/ml
Western Blot (WB)
(Figure 1. Western blot analysis of CARD4 using anti-CARD4 antibody (MBS177898).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: A549 Whole Cell Lysate,Lane 2: Rat Cardiac Muscle Tissue Lysate.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 antigen affinity purified polyclonal antibody at 0.5ug/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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for CARD4 at approximately 107KD. The expected band size for CARD4 is at 107KD. )
Western Blot (WB)
(Figure 1. Western blot analysis of CARD4 using anti-CARD4 antibody (MBS177898).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: A549 Whole Cell Lysate,Lane 2: Rat Cardiac Muscle Tissue Lysate.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 antigen affinity purified polyclonal antibody at 0.5ug/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 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for CARD4 at approximately 107KD. The expected band size for CARD4 is at 107KD. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of CARD4 using anti-CARD4 antibody (MBS177898).CARD4 was detected in paraffin-embedded section of Human Placenta Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-CARD4 Antibody (MBS177898) 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 2. IHC analysis of CARD4 using anti-CARD4 antibody (MBS177898).CARD4 was detected in paraffin-embedded section of Human Placenta Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-CARD4 Antibody (MBS177898) 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of CARD4 using anti-CARD4 antibody (MBS177898).CARD4 was detected in immunocytochemical section of A549 cell. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-CARD4 Antibody (MBS177898) 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 3. IHC analysis of CARD4 using anti-CARD4 antibody (MBS177898).CARD4 was detected in immunocytochemical section of A549 cell. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-CARD4 Antibody (MBS177898) 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of CARD4 using anti-CARD4 antibody (MBS177898).CARD4 was detected in immunocytochemical section of Hela cell. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-CARD4 Antibody (MBS177898) 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) with DAB as the chromogen. )
Immunohistochemistry (IHC)
(Figure 4. IHC analysis of CARD4 using anti-CARD4 antibody (MBS177898).CARD4 was detected in immunocytochemical section of Hela cell. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-CARD4 Antibody (MBS177898) 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) with DAB as the chromogen. )
Immunocytochemistry (ICC)
(Figure 5. IHC analysis of CARD4 using anti-CARD4 antibody (MBS177898).CARD4 was detected in immunocytochemical section of SMMC-7721 cell. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-CARD4 Antibody (MBS177898) 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) with DAB as the chromogen. )
Immunocytochemistry (ICC)
(Figure 5. IHC analysis of CARD4 using anti-CARD4 antibody (MBS177898).CARD4 was detected in immunocytochemical section of SMMC-7721 cell. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1ug/ml rabbit anti-CARD4 Antibody (MBS177898) 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) with DAB as the chromogen. )
Background: 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.
NCBI and Uniprot Product Information
NCBI Description
This gene encodes a member of the NOD (nucleotide-binding oligomerization domain) family. This member is a cytosolic protein. It contains an N-terminal caspase recruitment domain (CARD), a centrally located nucleotide-binding domain (NBD), and 10 tandem leucine-rich repeats (LRRs) in its C terminus. The CARD is involved in apoptotic signaling, LRRs participate in protein-protein interactions, and mutations in the NBD may affect the process of oligomerization and subsequent function of the LRR domain. This protein is an intracellular pattern-recognition receptor (PRR) that initiates inflammation in response to a subset of bacteria through the detection of bacterial diaminopimelic acid. Multiple alternatively spliced transcript variants differring in the 5' UTR have been described, but the full-length nature of these variants has not been determined. [provided by RefSeq, Oct 2009]
Uniprot Description
NOD1: Enhances caspase-9-mediated apoptosis. Induces NF-kappa- B activity via RIPK2 and IKK-gamma. Confers responsiveness to intracellular bacterial lipopolysaccharides (LPS). Forms an intracellular sensing system along with ARHGEF2 for the detection of microbial effectors during cell invasion by pathogens. Required for RHOA and RIPK2 dependent NF-kappa-B signaling pathways activation upon S.flexneri cell invasion. Involved not only in sensing peptidoglycan (PGN)-derived muropeptides but also in the activation of NF-kappa-B by Shigella effector proteins IpgB2 and OspB. Homodimer. Self-associates. Binds to caspase-9 and RIPK2 by CARD-CARD interaction. Interacts with ARHGEF2. Highly expressed in adult heart, skeletal muscle, pancreas, spleen and ovary. Also detected in placenta, lung, liver, kidney, thymus, testis, small intestine and colon.
Chromosomal Location of Human Ortholog: 7p14.3
Cellular Component: apical plasma membrane; basolateral plasma membrane; cytoplasm; cytosol
Molecular Function: ATP binding; CARD domain binding; caspase activator activity; identical protein binding; peptidoglycan binding; protein binding; protein homodimerization activity
Biological Process: activation of MAPK activity; activation of NF-kappaB transcription factor; apoptosis; caspase activation; defense response; defense response to bacterium; defense response to Gram-positive bacterium; detection of bacterium; detection of biotic stimulus; inflammatory response; innate immune response; interleukin-8 biosynthetic process; JNK cascade; positive regulation of caspase activity; positive regulation of dendritic cell antigen processing and presentation; positive regulation of I-kappaB kinase/NF-kappaB cascade; positive regulation of interleukin-1 beta production; positive regulation of interleukin-6 production; positive regulation of JNK cascade; positive regulation of nitric-oxide synthase activity; positive regulation of tumor necrosis factor production; protein oligomerization; signal transduction
Research Articles on CARD4
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Product Notes
The CARD4 nod1 (Catalog #AAA177898) is an Antibody and is intended for research purposes only. The product is available for immediate purchase. The Anti-CARD4 Antibody reacts with Human, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's CARD4 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry (IHC) Paraffin. Western Blot Concentration: 0.1-0.5ug/ml Immunohistochemistry (IHC) Paraffin Concentration: 0.5-1ug/ml. Researchers should empirically determine the suitability of the CARD4 nod1 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "CARD4, Polyclonal Antibody" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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