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Authors: Kelsey Miller
Group: BioLegend
Summary: The cells targeted by the Nanobeads are either selected or depleted by incubating your sample with the directly conjugated magnetic particles. The magnetically labeled fraction is retained by the use of a magnetic separator. After collection of the targeted cells, downstream applications include functional assays, gene expression, phenotypic characterization, etc.
Proper citation: Kelsey Miller 2016. MojoSort™ Nanobeads No Wash Protocol. protocols.io dx.doi.org/10.17504/protocols.io.e2tbgen Copy
Authors: Kelsey Miller
Group: BioLegend
Summary: This procedure is optimized for the isolation of 107 to 2 x 108 cells per tube. If working with fewer than 107 cells, keep volumes as indicated for 107 cells. For best results, optimize the conditions to your specific cell number and tissue. Prepare fresh MojoSort™ Buffer solution by diluting the 5X concentrate with sterile distilled water. The purity of the CD3- CD49b+ NK Cell fraction typically ranges from 80 to 90%. Removal of dead cells is highly recommended prior to cell isolation. Dead cells can be removed using appropriate centrifugation media or other methods.Product description and procedure summary: The kit is designed for the isolation of mouse NK cells from lymphoid tissues. Target cells are depleted by incubating the sample with the biotin antibody cocktail followed by incubation with magnetic Streptavidin Nanobeads. The magnetically labeled fraction is retained by the use of a magnetic separator. The untouched cells are collected. These are the cells of interest; do not discard the liquid. Some of the downstream applications include functional assays, gene expression, phenotypic characterization, etc.
Proper citation: Kelsey Miller 2016. MojoSort™ Mouse NK Cell Isolation Kit Protocol. protocols.io dx.doi.org/10.17504/protocols.io.fxpbpmn Copy
Authors: Kelsey Miller
Group: BioLegend
Summary: This protocol is essentially the same as a standard ELISA protocol, with one major exception – all animal compounds must be avoided in buffers, etc. because they contain Neu5Gc, which will interfere with antibody activity and result in misleading data. Essentially, the glycoconjugate to be evaluated is immobilized on an ELISA plate and exposed to a primary antibody. The antibody provided in this kit has been shown to identify as little as 5 pmol of Neu5Gc per microgram glycoprotein, which is at or below the current detection limit for conventional DMB HPLC analysis. An advantage of the ELISA procedure is that it confirms the presence of Neu5Gc on the glycoconjugate of interest, but use caution, as it also reacts with solution/media that contains glycoconjugate.
Proper citation: Kelsey Miller 2016. Anti-Neu5Gc Antibody Kit Protocol - ELISA. protocols.io dx.doi.org/10.17504/protocols.io.e27bghn Copy
Authors: Kelsey Miller
Group: BioLegend
Summary: Using UV-induced peptide exchange, MHC/peptide monomers can be generated with conditional Flex-T™ monomers that harbor peptides of interest in their binding grooves. These new MHC monomers are subsequently multimerized using streptavidin-fluorophore conjugates. The resulting Flex-T™ reagents can be used for staining antigen-specific T cells and flow cytometric analysis. In humans, the MHC molecules are called HLA (Human Leukocyte Antigen).
Proper citation: Kelsey Miller 2017. Flex-T™ Tetramer and Cell Staining Protocol. protocols.io dx.doi.org/10.17504/protocols.io.hv6b69e Copy
Authors: Kelsey Miller
Group: BioLegend
Proper citation: Kelsey Miller 2016. Immunohistochemistry Protocol for Keratin Antibodies. protocols.io dx.doi.org/10.17504/protocols.io.e2hbgb6 Copy
Authors: Kelsey Miller
Group: BioLegend
Summary: The Anti-Neu5Gc Antibody Kit contains the essential monospecific polyclonal chicken IgY antibody, along with a negative control primary antibody to detect the presence of Neu5Gc on glycoconjugates by Western blot (WB). Samples to be evaluated are first subjected to SDS-PAGE, followed by transfer to a nitrocellulose or polyvinylidenedifluoride (PVDF) membrane. The membrane is then incubated with affinity-purified polyclonal anti-Neu5Gc to determine the presence of Neu5Gc on the protein of interest.The antibody provided in this kit has been shown to identify as little as 5 pmol of Neu5Gc per ug glycoprotein, which is at or below the current detection limit for conventional analysis by acid release, purification, DMB derivatization, HPLC, and electrospray mass-spectrometry. The Western blot provides additional information in that it confirms that Neu5G is directly linked to the glycoprotein of interest rather than to an accompanying sample component.
Proper citation: Kelsey Miller 2017. Anti-Neu5Gc Antibody Kit Protocol - Western Blot. protocols.io dx.doi.org/10.17504/protocols.io.hvrb656 Copy
Authors: Kelsey Miller
Group: BioLegend
Summary: Anti-BrdU Staining Protocol Using 70% Ethanol and 2N HCl
Proper citation: Kelsey Miller 2016. Anti-BrdU Staining Using 70% Ethanol and 2N HCl. protocols.io dx.doi.org/10.17504/protocols.io.e2pbgdn Copy
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