Anti-CD226 Antibody Picoband™

Product Name

Anti-CD226 Antibody Picoband™

View all DNAM-1/CD226 Antibodies

SKU/Catalog Number

A01094-2

Size

100 μg/vial

Form

Lyophilized

Description

Boster Bio Anti-CD226 Antibody Picoband™ catalog # A01094-2. Tested in ELISA, Flow Cytometry, IHC, WB applications. This antibody reacts with Mouse, Rat.

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-CD226 Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01094-2)

Host

Rabbit

Contents

Each vial contains 4mg Trehalose, 0.9mg NaCl and 0.2mg Na2HPO4.

Clonality

Polyclonal

Isotype

Rabbit IgG

Immunogen

E.coli-derived mouse CD226 recombinant protein (Position: E31-L333).

*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

A01094-2 is reactive to CD226 in Mouse, Rat

Applications

A01094-2 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

70 kDa

Calculated molecular weight

38.614kDa

Background of DNAM-1/CD226

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.

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, Mouse, Rat
Immunohistochemistry (Paraffin-embedded Section), 2-5μg/ml, Mouse, Rat
Flow Cytometry, 1-3μg/1x10⁶ cells, Rat
Direct ELISA, 0.1-0.5μg/ml, Mouse

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of CD226 using anti-CD226 antibody (A01094-2).
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 30ug of sample under reducing conditions.
Lane 1: rat spleen tissue lysates,
Lane 2: mouse spleen tissue lysates,
Lane 3: mouse thymus tissue lysates,
Lane 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 # A01094-2) 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 CD226 at approximately 70KD. The expected band size for CD226 is at 70KD.

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Figure 2. IHC analysis of CD226 using anti-CD226 antibody (A01094-2).
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 (A01094-2) 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.

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Figure 3. IHC analysis of CD226 using anti-CD226 antibody (A01094-2).
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 (A01094-2) 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.

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Figure 4. Flow Cytometry analysis of NRK cells using anti-CD226 antibody (A01094-2).
Overlay histogram showing NRK cells stained with A01094-2 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-CD226 Antibody (A01094-2, 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.

Gene/Protein Information For CD226 (Source: Uniprot.org, NCBI)

Alternative Names

CD226 antigenplatelet and T cell activation antigen 1; CD226 molecule; CD226; DNAM1; DNAM-1; DNAM1adhesion glycoprotein; DNAM-1DNAX accessory molecule-1; DNAX accessory molecule 1; PTA1; T lineage-specific activation antigen 1 antigen; TLiSA1 CD226 DNAM-1, DNAM1, PTA1, TLiSA1 CD226 molecule CD226 antigen|DNAX accessory molecule-1|T lineage-specific activation antigen 1 antigen|adhesion glycoprotein|platelet and T cell activation antigen 1

For more info on CD226, check out the CD226 Infographic

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In this infographic, you will see the following information for CD226: 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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