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vTAG™ ANTI-HUMAN CD45 ANTIBODY
NO WASH, QUANTITATIVE CD45 MEASUREMENT BY vFC™

$45.00 – $135.00

Conjugate(s): PE
Est. Limit of Detection: 30 Molecules per EV

 

Cargo analysis using vTag™ antibodies should be accompanied by a vesicle specific analysis method such as vFC™. vTag™ antibodies bundled with the vFC™ assay will be discounted 25%.
Certificate of Analysis
TDS MSDS
SKU: CBS66 Category:

Description

Product Attributes

Tested Reactivity: Anti-Human.
Other Published Reactivity: .
Conjugate: PE (see dropdown at the top for available options).
Sensitivity: Limit of detection is estimated at 30 molecules per vesicle in vFC™ for the PE conjugate. Change selection above for LOD of the antibody conjugated to a different fluorophore. LOD will vary depending upon sample, instrument, and other experimental conditions and should be empirically determined for your system.
Application: Optimized for vFC™ with no wash steps required.
Clonality: Monoclonal.
Clone: 2D1.
Host and Isotype: Mouse IgG1, kappa.
Positive Control Sample: .
Localization of Staining: Vesicle Surface.
Buffer and Stabilizer: 10mM PBS with 0.05% BSA & 0.05% azide.
Concentration: 5 ul/test.
Purification Method: Affinity purified during antibody production, additionally purified during conjugation to remove free dye and unbound antibody..
Immunogen: Human PBMC.
Storage Conditions: Store undiluted between 2°C and 8°C. Protected from prolonged exposure to light. Do not freeze. Stable for 1 year when properly stored..

Protocols

CD9 Immunostaining in vFC™

vFC™ (#CBS4) is an assay that enables specific detection and characterization of individual extracellular vesicles such as exosomes and microvesicles on commercially available flow cytometers. This protocol makes use of vFC™ to provide a standardized means of analyzing individual vesicles to measure size, concentration, and CD45 expression simultaneously. Because vFC™ triggers on vesicles, pairing antibody staining with vFC™ enables a highly-reproducible, no-wash assay. In this protocol, vesicles will be stained using our vTag™ Anti-Human CD45 Antibody. See protocol 2 in the vFC™ kit handbook for additional considerations and full protocol details.Procedure

We recommend using vTag™ Antibodies with the vFC™ Assay. Full, detailed protocols are available on the product page.
  1. Dilute sample containing EVs to approximately 5×106 extracellular vesicles per µl. To empirically determine vesicle concentration and identify an optimal pre-stain dilution, see supporting protocols included in the vFC™ kit.
  2. Prepare a working concentration of vFRed™ Red (5 µl per well) according to kit instructions
  3. In duplicate, in the included 96 well plate, add 35 µl of dilution buffer, 5µl of vFRed™ Red, 5 µl of vTag™ anti-human CD45 antibody, and 5 µl of EV sample.
  4. Incubate in the dark for 1hr.
  5. Perform serial dilution of the samples as described in the assay handbook. Briefly, samples are to be diluted from between 1:100 to 1:1000 from staining concentration depending upon the flow cytometer being used.
  6. Analyze the diluted sample of labeled EVs on a qualified flow cytometer.

CD45 Background Information

CD45 (also known as Leukocyte Common Antigen) is a protein tyrosine phosphatase expressed on all nucleated hematopoietic cells, where it regulates signaling pathways involved in cell activation and immune responses. It is essential for lymphocyte development and function. (see the data by vesicle origin resource for more info).

Additional information

Size

20 Tests, 100 Tests
Conjugate

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NO WASH, QUANTITATIVE CD45 MEASUREMENT BY vFC™
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vTAG™ ANTI-HUMAN CD45 ANTIBODY
NO WASH, QUANTITATIVE CD45 MEASUREMENT BY vFC™
$45.00 – $135.00