Tissue Homogenization for Lactic Acid Level - (Apr/06/2007 )
Hi
I want to know the process of Skeletal Muscle Homogenization for the detection of lactic Acid Level please reply as early as possible
I want to know the process of Skeletal Muscle Homogenization for the detection of lactic Acid Level please reply as early as possible
You can homogenize it, then centrifuge to get the supernatant to measure lactic level
Lactic acid is usually measured from the blood. You just need to separate the cells from the rest of the gunk (i.e work with serum). Hungry cells will metabolize glucose to lactic acid which would give you a flase positive. Lactate leaks through the membrane anyway, though. If you need to use the muscle cells themslves, either chemical lysis (lysozyme) or mechanical would do. You could analyse the lactate in the fluid before lysis and again after to monitor the effectiveness of whatever method you use in comparison with the leakage effect. You'd need a control consisting of the cells left soaking in the fluid with no lytic intervention.
Sounds like fun.
Thank you for reply
the Muscle Homogenization process i need, You did'nt clearly mentioned the. Kindly send me the Procedure of Skeletal muscle homogenization for the Lactic acid level measurement.
I shall be really thankfull.
Please send the reference regarding homogenization of skeletal muscle for the lactic acid level
regards
dr khalid
Sounds like fun.
thanks for reply
Kindly send me the exact procedure and reference for the Lactic acid level detection in skeletal muscle and what is the proper way to homogenize the muscle
thanks
I want to know the process of Skeletal Muscle Homogenization for the detection of lactic Acid Level please reply as early as possible
You can homogenize it, then centrifuge to get the supernatant to measure lactic level
Two methods to homogenise: either freeze the tissue (immerse in a Dewar of dry ice) then immediatly crack with a hammer. We designed a metal well into which fitted a metal punch. We put our frozen tissue straight into the hole and cracked it with a hammer. The other method would be simply to put the tissue in a homogenizer (electrical "food mixer" type) or ye olde worlde mortar and pestle. Use the first if you need to STOP any reaction quickly (which is what we were doing). Alternativly, use lysozyme to punch holes in the membrane (4mg/ml). You could do a time course study on lactate released with either the homogenizer or lysozyme method. (Could be a student project?)
Best of luck