this protein marker standard should be used. Figure 1: SDS PAGE Gel of Murine Fi
ID: 162786 • Letter: T
Question
this protein marker standard should be used.
Figure 1: SDS PAGE Gel of Murine Fibroblast Cell Lysate samples. Lane 1, 5 µL protein marker added. Lane 2, sample 3 containing 5 µL of a 16 µL of cell lysate solution. Lane 3, containing of 5 µL of a 9.6 µL cell lysate and 6.4 µL distilled water dilution solution. Lane 5, containing 5 µL of an 8 µL Bovine Serum and 32 µL distilled water solution. Lane 6, containing 5 µL of a 4 µL Bovine Serum and 16 µL of distilled water solution. Lane 7, containing 5 µL of a 2 µL Bovine Serum and 8 µL of distilled water solution. Lane 8, containing 5 µL of an 8 µL Horse Serum and 32 µL distilled water solution. Lane 9, containing 5 µL of a 4 µL Horse Serum and 16 µL of distilled water solution. Lane 10, containing 5 µL of a 2 µL Horse Serum and 8 µL of distilled water solution.
Fro left to right: Lane 10, Lane 9, Lane 8, Lane 7, Lane 6, Lane 5, Lane 4, Lane 3, Lane 2. Lane 1
QUESTIONS:
Anaylze two protein bands out of lanes 5-7, two protein bands out of lanes 8-10 & lane 2 and 3 bands.
examples on how to anaylze:
1. Size, clarity (is it fuzzy, straight, etc).
2. Estimate molecular masses and relative abundance of unknown polypeptides in a complex mixture
3. Patterns of bands that suggest presence of proteins
4. Effectiveness of a separation procedure
5. Compare and contrats on band to another
Then:
4. Guess what proteins are in the serum shown in the gel result
5. Discuss and critique gel quality:Overloading, under-loading, distortion of lanes, non-reproducible patterns, smears, streaks, etc.
Explanation / Answer
bands out of lanes 5-7
The protein loaded (Bovine serum) in the lane 5 is looking more than the other lane but the dilution and the amount is same in all the lane (5-7) this is indicating the unequal loading.
Because of the more amount of the protein the bands are distorted so the amount of the protein loaded should be reduced.
Out of the three band the banding pattern and sharpness of the band for lane 7 is good but amount should be reduced for better comparision.
bands out of lanes 8-10
The protein (Horse serum) loaded in the lane 8 is looking more than the other lane but the dilution and the amount is same in all the lane (8-10) this is indicating the unequal loading.
Because of the more amount of the protein the bands are distorted so the amount of the protein loaded should be reduced.
Out of the three band the banding pattern and sharpness of the band for lane 10 is good but amount should be reduced for better comparision.
bands out of lanes 2-3
The lysate loaded in the gel is very less we are hardly able to seethe band in the 2nd lane and 3rd lane nothing is visible.
The bands are not sharp may be becuase of the high salt and detergent in the lysate that should be diluted little bit to reduce the salt and detergent concentration to get a sharp band.
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