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Western Blot (WB) (Figure 1. Western blot analysis of CD226 using anti-CD226 antibody (MBS1753396).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 30ug of sample under reducing conditions.Lane 1: rat spleen tissue lysatesLane 2: mouse spleen tissue lysatesLane 3: mouse thymus tissue lysatesLane 4: mouse Raw264. 7 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-CD226 antigen affinity purified polyclonal antibody (Catalog # MBS1753396) 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 CD226 at approximately 70KD. The expected band size for CD226 is at 70KD. )

Rabbit anti-Mouse, Rat CD226 Polyclonal Antibody | anti-Cd226 antibody

Anti-CD226 Antibody

Gene Names
Cd226; Pta1; DNAM1; DNAM-1; TLiSA1; BC051526
Reactivity
Mouse, Rat
Applications
Western Blot, Immunohistochemistry, Flow Cytometry, Functional Assay, ELISA
Purity
Immunogen affinity purified.
Synonyms
CD226; Polyclonal Antibody; Anti-CD226 Antibody; Cd226; Pta1; CD226 antigen; Platelet and T-cell activation antigen 1; CD antigen CD226; anti-Cd226 antibody
Ordering
For Research Use Only!
Host
Rabbit
Reactivity
Mouse, Rat
Clonality
Polyclonal
Isotype
Rabbit IgG
Specificity
Rabbit IgG polyclonal antibody for CD226 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-Cd226 antibody
Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA)
Application Notes
WB: 0.25-0.5ug/ml|Mouse, Rat|
IHC-P: 2-5ug/ml|Mouse, Rat|
FC/FACS/FCM: 1-3ug/1x106 cells|Rat|
Direct ELISA: 0.1-0.5ug/ml|Mouse|
Immunogen
E Coli-derived mouse CD226 recombinant protein (Position: E31-L333).
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.

Western Blot (WB)

(Figure 1. Western blot analysis of CD226 using anti-CD226 antibody (MBS1753396).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 30ug of sample under reducing conditions.Lane 1: rat spleen tissue lysatesLane 2: mouse spleen tissue lysatesLane 3: mouse thymus tissue lysatesLane 4: mouse Raw264. 7 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-CD226 antigen affinity purified polyclonal antibody (Catalog # MBS1753396) 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 CD226 at approximately 70KD. The expected band size for CD226 is at 70KD. )

Western Blot (WB) (Figure 1. Western blot analysis of CD226 using anti-CD226 antibody (MBS1753396).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 30ug of sample under reducing conditions.Lane 1: rat spleen tissue lysatesLane 2: mouse spleen tissue lysatesLane 3: mouse thymus tissue lysatesLane 4: mouse Raw264. 7 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-CD226 antigen affinity purified polyclonal antibody (Catalog # MBS1753396) 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 CD226 at approximately 70KD. The expected band size for CD226 is at 70KD. )

Immunohistochemistry (IHC)

(Figure 2. IHC analysis of CD226 using anti-CD226 antibody (MBS1753396).CD226 was detected in paraffin-embedded section of mouse lung 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-CD226 Antibody (MBS1753396) 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. )

Immunohistochemistry (IHC) (Figure 2. IHC analysis of CD226 using anti-CD226 antibody (MBS1753396).CD226 was detected in paraffin-embedded section of mouse lung 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-CD226 Antibody (MBS1753396) 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. )

Immunohistochemistry (IHC)

(Figure 3. IHC analysis of CD226 using anti-CD226 antibody (MBS1753396).CD226 was detected in paraffin-embedded section of rat lung lymphaden 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-CD226 Antibody (MBS1753396) 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. )

Immunohistochemistry (IHC) (Figure 3. IHC analysis of CD226 using anti-CD226 antibody (MBS1753396).CD226 was detected in paraffin-embedded section of rat lung lymphaden 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-CD226 Antibody (MBS1753396) 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. )

Flow Cytometry (FC/FACS)

(Figure 4. Flow Cytometry analysis of NRK cells using anti-CD226 antibody (MBS1753396).Overlay histogram showing NRK cells stained with MBS1753396 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CD226 Antibody (MBS1753396, 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. )

Flow Cytometry (FC/FACS) (Figure 4. Flow Cytometry analysis of NRK cells using anti-CD226 antibody (MBS1753396).Overlay histogram showing NRK cells stained with MBS1753396 (Blue line). The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CD226 Antibody (MBS1753396, 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. )
Related Product Information for anti-Cd226 antibody
DNAM 1, also known as CD226, is a receptor expressed by peripheral blood T lymphocytes that is involved in intercellular adhesion, lymphocyte signaling, cytotoxicity and lymphokine secretion mediated by cytotoxic T lymphocytes. DNAM 1 is broadly expressed on T cells, NK cells, platelets, monocytes and a subset of B cells. DNAM-1 is also expressed by a subset of CD3 positive thymocytes. This antibody is reported to inhibit T- and NK cell mediated cytotoxicity against tumour cell targets and to block TNF alpha and IFN gamma secretion by alloantigen-specific T-cells.
References
1. Bottino, C., Castriconi, R., Pende, D., Rivera, P., Nanni, M., Carnemolla, B., Cantoni, C., Grassi, J., Marcenaro, S., Reymond, N., Vitale, M., Moretta, L., Lopez, M., Moretta, A. Identification of PVR (CD155) and nectin-2 (CD112) as cell surface ligands for the human DNAM-1 (CD226) activating molecule. J. Exp. Med. 198: 557-567, 2003.
2. Iguchi-Manaka, A., Kai, H., Yamashita, Y., Shibata, K., Tahara-Hanaoka, S., Honda, S., Yasui, T., Kikutani, H., Shibuya, K., Shibuya, A. Accelerated tumor growth in mice deficient in DNAM-1 receptor. J. Exp. Med. 205: 2959-2964, 2008.
3. Liu, J., Qian, X., Chen, Z., Xu, X., Gao, F., Zhang, S., Zhang, R., Qi, J., Gao, G. F., Yan, J. Crystal structure of cell adhesion molecule nectin-2/CD112 and its binding to immune receptor DNAM-1/CD226. J Immun. 188: 5511-5520, 2012.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
38,063 Da
NCBI Official Full Name
CD226 antigen isoform a
NCBI Official Synonym Full Names
CD226 antigen
NCBI Official Symbol
Cd226
NCBI Official Synonym Symbols
Pta1; DNAM1; DNAM-1; TLiSA1; BC051526
NCBI Protein Information
CD226 antigen; platelet and T cell activation antigen 1; platelet and T-cell activation antigen 1
UniProt Protein Name
CD226 antigen
Protein Family
UniProt Gene Name
Cd226
UniProt Synonym Gene Names
Pta1
UniProt Entry Name
CD226_MOUSE

Uniprot Description

DNAM-1: Receptor involved in intercellular adhesion, lymphocyte signaling, cytotoxicity and lymphokine secretion mediated by cytotoxic T-lymphocyte (CTL) and NK cell.

Protein type: Immunoglobulin superfamily; Membrane protein, integral; Cell adhesion

Cellular Component: cell surface; membrane; integral to membrane; external side of plasma membrane

Molecular Function: integrin binding; protein binding; cell adhesion molecule binding; protein kinase binding

Biological Process: positive regulation of natural killer cell mediated cytotoxicity directed against tumor cell target; positive regulation of immunoglobulin mediated immune response; cytokine production; positive regulation of natural killer cell mediated cytotoxicity; cell adhesion; positive regulation of mast cell activation

Research Articles on Cd226

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Product Notes

The Cd226 cd226 (Catalog #AAA1753396) is an Antibody produced from Rabbit and is intended for research purposes only. The product is available for immediate purchase. The Anti-CD226 Antibody reacts with Mouse, Rat and may cross-react with other species as described in the data sheet. AAA Biotech's CD226 can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB), Immunohistochemistry-Paraffin (IHC-P), Flow Cytometry (FC/FACS/FCM), Direct ELISA (EIA). WB: 0.25-0.5ug/ml|Mouse, Rat| IHC-P: 2-5ug/ml|Mouse, Rat| FC/FACS/FCM: 1-3ug/1x106 cells|Rat| Direct ELISA: 0.1-0.5ug/ml|Mouse|. Researchers should empirically determine the suitability of the Cd226 cd226 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 "CD226, 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.

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