Immunohistochemistry Wilms Tumor 1 (WT1) Rabbit Mouse

Get tips on using ON-TARGETplus Mouse Rheb (19744) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - RAW264.7 Rheb

The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.

Get tips on using ON-TARGETplus Mouse Fdps (110196) siRNA - Individual, to perform siRNA / miRNA gene silencing Mouse - MC3T3-E1 Fdps

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Get tips on using ON-TARGETplus Mouse Myb (17863) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - NIH-3T3 Myb

Get tips on using ON-TARGETplus Mouse Eif2ak3 (13666) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - CT26 Perk/Eif2ak3

Get tips on using ON-TARGETplus Mouse Casp8 (12370) siRNA - SMARTpool to perform siRNA / miRNA gene silencing Mouse - CT26 caspase-8

Get tips on using Ni-NTA Magnetic Agarose Beads (6 x 1 ml) to perform Protein tag Purification of His-tagged proteins

An alternative to culture-based cell death detection is an assessment of other cell viability indicators using fluorescent dyes, including membrane potential and membrane integrity. Live/Dead assays differentiates live and dead cells using membrane integrity as a proxy for cell viability and are based on a fluorescent staining procedure followed by detection using flow cytometry. However, samples preparation for such flow cytometry-based techniques could be challenging. Cell harvesting by trypsinization, mechanical or enzymatic cell disaggregation from tissues, extensive centrifugation steps, may all lead to preferential loss of apoptotic cells. To overcome this strictly follow manufacturers instruction of the detection kit.

Cell Invasion or Cell Migration assays are technically challenging to set up as the gradient between the two compartments equilibrates in time during the assay. It is also problematic to view cells and for cells to migrate through a non-physiologic polycarbonate or polypropylene filter. Care must be taken while loading the well with cells to form a single cell suspension. Precaution must be taken while trypsinization (under-trypsinization can lead to cell clumping while over-trypsinization could strip off adhesion molecules necessary for migration). This leads to difficulty in getting significant results, when only small numbers of cells cross the filter or when the distribution and/or staining of the cells is uneven.