![]() The Xenopus oocyte: A single-cell model for studying Ca 2+ signaling. Nuclear microinjection to assess how heterologously expressed proteins impact Ca 2+ signals in Xenopus oocytes. Unique protein kinase C profile in mouse oocytes: lack of calcium-dependent conventional isoforms suggested by rtPCR and Western blotting. The biochemical basis of an all-or-none cell fate switch in Xenopus oocytes. Regulation of mouse egg activation: Presence of ryanodine receptors and effects of microinjected ryanodine and cyclic ADP ribose on uninseminated and inseminated eggs. Reviewed DecemWestern blotting is a technique that uses specific antibodies to identify proteins that have been separated based on size by gel electrophoresis. Western blot analysis indicated that there is also an overall increase in ser 23/24 phosphorylation in rAMPK/LKB1 hi-treated fibers. In one protocol, 20 nM of LKB1/Mo25/STRAD (LKB1 complex Millipore. The procedure is faster with the Trans-Blot Turbo system (time for electrophoresis and transfer is twice as short) and is more amenable to higher throughput (twice the number of gels can be processed simultaneously) such that the entire procedure is complete in ~1 h. Our western blot protocol includes solutions and reagents, procedures, and useful links to guide you through your experiment. Separate protocols were used to activate the AMPK complex. Both these systems are convenient, easy to use, and show single-cell sensitivity for detecting the transfer of large, endogenous proteins (e.g., Xenopus type 1 InsP 3 receptor). Different commercial systems are available, for example iBlot and Trans-Blot Turbo, and both have proved suitable for this purpose, although selection of compatible consumables is important. The recent introduction of rapid western transfer equipment has streamlined procedures for analyzing oocytes used for functional assays (e.g., Ca 2+ imaging). 1) and the importance of such single cell correlations of expression/activity are well evidenced relative to population analyses ( Ferrell and Machleder 1998). Immunodetection of expressed gene products in Xenopus oocytes is easily performed at the single cell and/or regional level ( Fig. This concern is obviously less acute in the Xenopus system given the large quantity of protein in a single oocyte (125 μg), and the availability of oocytes (~15% mass of a frog). For my western blot protocol, I searched what method to follow and this were the steps that I followed. For certain intracellular Ca 2+ channel isoforms, extracts from as many as 5000 cells have been needed for immunoprecipitations and western blotting analyses ( Ayabe et al. To detect relatively scarce proteins in mouse oocytes, hundreds of oocytes may be needed for a single lane immunodetection, and a single mouse contains only tens of oocytes ( Gangeswaran and Jones 1997). These premium antibodies are lab-validated using strict testing criteria to ensure superior performance in western blotting detection.In many animal models, running a western blot from oocytes/eggs can be challenging. PrecisionAb Validated Western Blotting Antibodies.SureBeads Protein G Magnetic Beads enable fast, easy, consistent immunoprecipitation without centrifugation. Loading buffers for protein or nucleic acid samples, detergents, ampholytes, and gel-casting buffers. For any inquiry or request for repair service, contact Bio-Rad Laboratories after con- firming the model and serial number of your. Protein Electrophoresis Buffers and Reagents.Interpretation of western-blot bands for CXCL16 was according to Gough et al. ![]() Our electrophoresis chambers enable rapid, high-resolution protein separation on precast or handcast gels over a variety of different gel sizes. Densitometric analysis was performed with Quantity One software (Biorad). Protocol Document Click here for example data file. Blot Absorbent Filter Paper (extra thick) Recommended uses 170-3966 Sheets, 7 x 8.4 cm pre-cut to fit mini PROTEAN 3 Gels, 60 170-3967 Sheets, 8.3 x 13.6 cm pre-cut to fit. Choose SDS-PAGE and native PAGE gels, convert to TGX Precast Gels, or choose specialized gel chemistries. western blot wet transfer bio rad, Western Blot Wet Transfer Sino, Mini Transblot Systems (for mini and maxi gels, respectively) Biocompare Product Review. Section 6 Examples of Specific Protocols. Prestained and unstained molecular weight standards for protein electrophoresis applications including SDS-PAGE, western blotting, 2-D PAGE, and isoelectric focusing (IEF).įind the right Bio-Rad protein gel for your application. Protein Ladders and Standards (Markers). ![]()
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