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#1
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| Dear all, i recently extracted proteins from a plant leaf, root and shoot and intended to analyze the proteins by SDS PAGE. i prepared these samples by crushing the sample( i.e root, shoot, leaf) in extraction buffer, sterile distilled water and sample buffer. But i think i over diluted the samples. Once i ran SDS PAGE with these samples ( the maximum sample i can load is 30micro l)...i observed that the bands were tooo faint...and also there was a lot of trailing in lanes. Could any 1 suggest ways to concentrate this already prepared sample so that i can get clear and distinct bands pleasee. THNX NIS |
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#2
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| U can precipitate your proteins using common TCA/acetone precipitation protocol as below: 1. Add 1 volume of 100% TCA to 4 volumes of protein sample. 2. Incubate 10 min at 4°C. 3. Centrifuge at 14K rpm, 4°C for 10mins. 4. Remove supernatant and wash pellet with 200μl cold acetone. 5. Centrifuge at 14K rpm,4°C for 5mins. 6. Repeat steps 4-5 once again. 7. Air dry pellet and resuspend in appropriate buffer. |
| The Following User Says Thank You to JennYeap For This Useful Post: | ||
admin (08-06-2011)
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| page , sample , sds , sds page |
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