nuclear Protocols

Category: Microscopy Protocols: Confocal Microscopy Protocols

Protocol describes a useful way to observe the development of embryos, as well as meristems & young primordia developing at the shoot apex by confocal microscopy after staining the nuclei with propidium iodide. The number of cells can be exactly quantified in a meristem or in young primordia. Because embryonic & meristematic cells are largely filled out by their nuclei, it is easier to image only the nuclei. This method allows analysis of whole-mount material, which is more easily reconstructed. - [Read Protocol for Nuclear Staining of Plants for Confocal Microscopy]

Category: Developmental Biology: Embryology Protocols

Protocol to Count Cell Number of Preimplantation Embryos using Nuclear Staining with Hoechst 33342 or DAPI. Includes: Preparation of Embryos; Preparation of Hoechst 33342 dye; Preparation of DAPI; Staining the Embryo; Mounting Embryos to Slides; What to Do When There are Too Many Cells to Count. - [Read Protocol to Count Cell Number of Preimplantation Embryos]

Category: Cell Culture Protocols: Insect Cell Culture Protocols

Includes protocols: Direct Adaptation to HyQ© SFX-Insect™ Medium; Sequential Adaptation to HyQ© SFX-Insect™ Medium; Adaptation of High Five™ Cells to HyQ© SFX-Insect™; Production of Recombinant Protein Using the Baculovirus Expression Vector System; Culturing Insect Cells and Production of Baculovirus Expressed Proteins in 2.8 L Fernbach Systems. Production of Autographa californica Multiple Capsid Nuclear Polyhedrosis Virus (rAcMNPV) Stock - [Read Protocols for Insect Cell Culture]

Category: Chromatography Protocols

Purification of Nuclear Factor DNA Recognition Site Affinity. PDF. - [Read Purification of Nuclear Factor DNA Recognition Site Affinity PDF]

Category: Cell Biology and Cell Culture

Protocol for the isolation of the lipid-rich microdomains of the plasma membrane, notably caveolae and lipid rafts. Methods for the isolation of lipid rafts are based on the insolubility of these structures in the nonionic detergent TritonX-100. Either the intact cells are treated with a detergent-containing solution or a post-nuclear supernatant is prepared from a cell homogenate and then Triton X-100 is added to this supernatant. - [Read S20 Purification of detergent-insoluble lipid rafts from cells and tissues.]

Category: Plant Biology Protocols: Plant Proteins, Peptides and Antigens

GUS is used as a tag to address nuclear localization whereas GFP is more versatile. GFP is detectable directly in living cells, GUS is only detected indirectly by staining of fixed tissue which may lead to artifacts or may obscure problems with protein solubility. In this protocol, protein localization is routinely assayed after particle-mediated transient transformation of onion epidermal cells. With this method it can be determined rapidly whether a given fusion protein is active and.... - [Read Subcellular Localization of GUS- and GFP-Tagged Proteins in Onion Epidermal Cells]

Category: Western Blot Protocols

Procedure describes how it denatures most of the modification and degradation proteins immediately giving the most accurate read out of the true levels of protein at the time of harvest.  However, in cases where detection is a problem, a limited purification (e.g. isolation of nuclear extract for the detection of transcription factors) might be required to allow analysis. - [Read Western Blot Analysis of Endogenous Gene Expression Protocol]

Category: Cell Organelle Protocols: Subcellular Fractionation Protocols

This protocol describes nuclear and cytoplasmic fractionation of tissue culture cells and a method for Western blot detection of proteins using the Odyssey Infrared Imaging System. This protocol was used to detect expression of the "small" Tap protein in 293T, HeLa and COS cells. The Odyssey system has several advantages over the more widely used chemiluminescent detection methods. - [Read Western Blot Analysis of Sub-Cellular Fractionated Samples Using the Odyssey Infrared Imaging System]

Category: Protein Protocols: Nuclear Extraction Protocols

Nuclear Extract preparation protocols, Buffers and solutions for nuclear extracts. Hahn Lab - [Read Yeast Nuclear Extract (Small Scale)]

Category: Yeast Protocols: Yeast Staining Protocols

Protocol describes the staining of nuclear and mitochondrial DNA with DAPI (4',6-diamidino-2-phenylindole). - [Read Yeast Vital Stains: DAPI Stain of Nuclear and Mitochondrial DNA Protocol]