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vTAG™ ANTI-HUMAN CD49F ANTIBODY
NO WASH, QUANTITATIVE CD49F MEASUREMENT BY vFC™

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

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SKU: CBS67 Category:

Description

Product Attributes

Tested Reactivity: Anti-Human.
Other Published Reactivity: African Green, Baboon, Capuchin Monkey, Cat, Cow, Chimpanzee, Cynomolgus, Dog, Horse, Rabbit, Sheep, Pig.
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: GOH3.
Host and Isotype: Rat IgG2a, 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: Mouse mammary tumor cells.
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 CD49F 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 CD49F 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 CD49F 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.

CD49F Background Information

CD49F (also known as Integrin α6) is a component of integrin complexes that includes α6β1 and α6β4 integrins. It is involved in cell adhesion, migration, and signaling by interacting with extracellular matrix proteins like laminin. CD49F plays a role in maintaining cell polarity, tissue architecture, and stem cell 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 CD49F MEASUREMENT BY vFC™
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