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Assay Kits & ELISAs

oYo-Link[R] MNase Antibody Labeling Reagent - Labels 100ug

Product Size(s)
Labels 100ug of antibody (50 CUT&RUN Reactions)
£529.00
AT6001-100
Academic Pricing
Please contact us for academic pricing.
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About this Product
SKU:
AT6001-100
Buffer:
10mM Tris-HCI (pH7.5), 50mM NaCl, 1mM EDTA, 50% Glycerol
Conjugate:
MNase
Extra Details:
oYo-Link[R] MNase allows for the site-specific labeling of All oYo-Link[R]-compatible antibodies with Micrococcal nuclease (MNase). MNase is derived from Staphylococcus aureus and is a relatively non-specific endonuclease that is capable of digesting single-stranded, double-stranded, circular and linear nucleic acids (DNA and RNA). MNase is widely used to study chromatin structure in chromatin immunoprecipitation (ChIP) assays by preferentially digesting naked DNA between nucleosomes and thus enriching the nucleosome- protected DNA fragments. The resulting undigested DNA is subjected to high-throughput sequencing and mapped to a reference genome. These assays can sometimes benefit from the direct conjugation of MNase to antibodies that target a desired transcription factor. Direct conjugation can also allow for epigenetic mapping within just a small number of cells, since the MNase can be directed to specific transcription factors or chromatin-associated proteins. One such approach is called the CUT&RUN Assay, whereby DNA is digested directly in cells using protein A (pA)-MNase or protein G (pG)-MNase conjugates. Since oYo-Link[R] MNase allows for the site-specific and covalent attachment of MNase directly to any antibody of choice, it can simplify in cell digestion assays by eliminating time-consuming incubation and wash-steps. It also prevents release of MNase. This can result in higher yield and higher resolution maps of genome-wide epigenetic changes. Requirements: To label your compatible antibodies with oYo-Link[R], you will need a Light Source emitting at 365nm (6-10W). Use your own Compatible Device from our list or purchase AlphaThera's LED Photo-Crosslinking Device. Technical Support: For additional questions concerning compatibility email support@alphathera.com
Shipping Conditions:
Dry Ice
Storage Conditions:
-20[o]C Freezer
Supplier:
AlphaThera
Size:
Labels 100ug of antibody (50 CUT&RUN Reactions)
Will oYo-Link work with my antibody?:
oYo-Link mIgG1 can be used to label mouse IgG1 antibody. Currently, oYo-Link mIgG1 is only available for certain products. Please check the product webpage. oYo-Link mIgG1 only labels mouse IgG1 and is not compatible with other antibodies.
Can oYo-Link be used to label antibodies in the presence of albumin and/or Tris?:
Yes, since oYo-Link specifically bind to the heavy chain of IgG, it works in the presence of both albumin and Tris. oYo-Link is compatible with all common buffers.
If an oYo-Link product has accidentally been left at room temperature for a week, will it still work?:
With the exception of oYo-Link MNase and oYo-Link DBCO, all oYo-Link products are shipped as white or clear pellets and are very stable. While we recommend cold storage, they will not exhibit any loss of activity if left at room temperature for a week. oYo-Link MNase and oYo-Link DBCO must be shipped and stored cold (-20°C) immediately upon arrival, otherwise there will be a loss in activity.
Does AlphaThera offer conjugation service?:
For custom conjugation services, you will need to send us your antibody. The conjugation and purification fee depends on the size and specifications of each order. Purification is not required for most applications.
How many oYo-Link labels will be conjugated to each antibody?:
For most subclasses and species of antibody, oYo-Link will result in the conjugation of 2 labels per antibody (maximum), one label per heavy chain. For example, ~95% of human IgG and Rabbit IgG will have two labels per antibody. However, there are a few antibodies such as mouse IgG2b and goat IgG, where the conjugation is slightly less efficient. In this case, 60 to 80% of the antibody will be labelled and have a mixture of 0, 1 and 2 labels per antibody.
How do I know if oYo-Link was successfully conjugated to my antibody?:
Antibody conjugation can be checked on SDS-PAGE gel. An example is present in the supporting data on each product page.
Does 365nm UV illumination damage the antibody and its binding affinity?:
The conjugated and unconjugated antibody binding affinities have been tested via both ELISA and cell binding assays and don’t show any difference.
Do I need to remove free oYo-Link after conjugating to antibody?:
For most immunoassay applications, purification is not necessary. In our standard recommended protocol, the molar ratio of oYo-Link-to-antibody is 5:1. These conditions ensure that the antibody conjugation efficiency reaches its maximum; however there will be a slight excess remaining in your antibody mixture. This is usually removed during immunoassay washing steps. However, if your application requires high antibody-conjugate purity, please contact us for more information on this process and for a detailed protocol.
For oYo-Link click-chemistry products, should I perform the click reaction before conjugation?:
When working with oYo-Link Azide (Catalog #: AT3002) and oYo-Link DBCO (Catalog #: AT3003), it is required to perform the click chemistry coupling reactions with the molecule of interest first, prior to photo-crosslinking to an antibody. This is to avoid UV illumination damage to the click moiety and to avoid diluting oYo-Link prior to the reaction to maximize the reaction efficiency. For oYo-Link Tetrazine (Catalog #: AT3004) and oYo-Link Thiol (Catalog #: AT3001); it is recommended to perform the coupling reaction before photo-crosslinking. This is to avoid diluting oYo-Link prior to the coupling reaction to maximize the reaction efficiency.
What is the lowest amount of antibody that can be labeled by oYo-Link reagents?:
We recommend working with at least 1 µg of antibody; however, it is possible that even lower amounts of antibody can be labelled.
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