Caution: Use caution while inserting the gel knife between the two plates to avoid excessive pressure towards the gel. Repeat on each side of the cassette until the plates are completely separated. Push down gently on the knife handle to separate the plates.The notched (“well”) side of the cassette should face up. Separate each of the three bonded sides of the cassette by inserting the Gel Knife into the gap between the cassette’s two plates.After electrophoresis is complete, shut off the power, disconnect electrodes, and remove gel(s) from the XCell SureLock™ Mini-Cell.*Expected start and end current values are stated for single gels. Run your gels according to the following protocol: See below for Electrophoresis Conditions. With the power on the power supply turned off, connect the electrode cords to the power supply. Place the XCell SureLock™ Mini-Cell lid on the Buffer Core.Fill the Lower Buffer Chamber with 600 ml of the appropriate 1X running buffer.Load appropriate protein molecular weight markers.Load an appropriate volume of sample at the desired protein concentration onto the gel (see Recommended loading volumes).The buffer level must exceed the level of the wells. Once the seal is tight, fill the Upper Buffer Chamber (inner) with the appropriate 1X running buffer.If you detect a leak from Upper to the Lower Buffer Chamber, discard the buffer, reseal the chamber, and refill. Fill the Upper Buffer Chamber with a small amount of the running buffer to check for tightness of seal.Note: If you are using only one gel, the plastic Buffer Dam replaces the second gel cassette. Refer to the XCell SureLock™ Mini-Cell manual (IM-9003) for detailed instructions. Seat the gels on the bottom of the Mini-Cell and lock into place with the Gel Tension Wedge. Orient the two gels in the Mini-Cell such that the notched “well” side of the cassette faces inwards toward the Buffer Core.Invert the gel and shake the gel to remove the buffer. Rinse the sample wells with the appropriate 1X SDS Running Buffer.In one smooth motion, gently pull the comb out of the cassette.Peel off the tape from the bottom of the cassette. Rinse the gel cassette with deionized water.Remove the Novex® Pre-Cast Gel from the pouch.XCell SureLock™ Mini-Cell requires 200 ml for the Upper Buffer Chamber and 600 ml for the Lower Buffer Chamber. Wear gloves and safety glasses when handling gels. If you are using any other mini-cell for electrophoresis, refer to the manufacturer’s recommendations. You may download this manual from our website or contact Technical Service. For more information on the XCell SureLock™ Mini-Cell, refer to the manual (IM-9003). Instructions are provided below for electrophoresis of the Novex® Pre-Cast Gels using the XCell SureLock™ Mini-Cell. Warning: This product contains a chemical (acrylamide) known to the state of California to cause cancer. Wear gloves at all times when handling gels. The Packaging Buffer contains 0.02% sodium azide and residual acylamide monomer. Gels are individually packaged in clear pouches with 4-10 ml of Packaging Buffer. The Novex® Pre-Cast Gels are supplied as 10 gels per box. Extended exposure of the gels to room temperature seriously impairs the performance of the gel. Use gels immediately from the refrigerator. The gels have a shelf life of 4-8 weeks depending upon the gel type when stored at +4° C. ![]() Recipes are provided on Recipes section if you are preparing your own buffers. During electrophoresis, the gel and buffer ions in the Tris-Glycine system form an operating pH of 9.5 in the separation region of the gel. Tris Base (+) is the common ion present in the gel buffer and running buffer.The running buffer ions are Tris+, Gly-, and dodecylsulfate- (pH 8.3). Glycine is partially negatively charged and trails behind the highly charged chloride ions in the charged environment. Glycine (-) is the primary anion supplied by the running buffer and serves as a trailing ion.The gel buffer ions are Tris+ and Cl- (pH 8.65). Chloride (-) is supplied by the gel buffer and serves as a leading ion due to its high affinity to the anode as compared to other anions in the system.The Tris-Glycine discontinuous buffer systems involves three ions: The separating range of Tris-Glycine gels is 6-200 kDa. The separating and stacking gels of Novex® Tris-Glycine gels have a pH of 8.65 unlike traditional Laemmli gels that have a stacking gel pH of 6.8 and separating gel pH of 8.8. The Tris-Glycine gels are based on the Laemmli System (Laemmli, 1970) with minor modifications for maximum performance in the pre-cast format.
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