The red arrow indicates where I am. I am trying to calculate the nanograms of pr

Question

The red arrow indicates where I am. I am trying to calculate the nanograms of protein per cell. The calculated cell density is 1.9 x 105 cells/mL. I calculated the protein per mL as 206.4 ug/mL since it was a 1:4 dilution (dont know if I did that right). I centrifuged 10 mL of the stock and multiplied that by the cell density to get 1.9 x 106 cells in the pellet. How would I calculate the ng protein per cell?

[Chart 1] Chart of the cell counts in A quadrants for calculating cell density Average cell count: (7+2+3+7)/4 = 9.5 Concentration (9.5) (2)/ (0.0001 mL)-1.9 x 105 cells/mL Graph 2] Graph of standard curve of Absorbance vs. Concentration of prgtein standards. Table 4] Absorbance results from the spectrophotometer readings on the protein standards Absorbance vs. Concentration of Protein Standarc Concentration 0.8 v-0.0081x +0.4173 0.9802 0.385 0564 078 0931 1.101 1.161 0.4 20 40 60 80 100 100 120 Concentration (ug/ml) [Table 5] Table of the absorbance readings from the dilution series lab 5 procedure the cell lysate outlined in step 5 of the Slope equation from Graph 2] used to find the diluted concentration of the 1:4 dilution. Y=0.0074x + 0.456 Dilution Absorbance 1:2 1:4 1:8 1:16 1.08 0.835 0.665 0.553 [1:4 dilution]: 0.835-0.0081x + 0.4173 [14] = 51.6 ug/mL Protein per mL: 51.6x 4 206.4 ug/mL 10 mL x 1.9 x 102 cells/mL 1.9 x 10° cells in the pellet since 10mL of culture was centrifuged

Explanation / Answer

The calculation that you have done to measure the protein concentration is correct.

Protein per ml = 206.4microgram/ml

Cell density is 1.9 *10^5 cells/ml.

If you have used the cell pellet of 10ml to make the protein lysate then you need to divide protein/ml with total cells.

206.4 / 1.9*10^6

1microgram = 1000 ng

So protein concentration will be 206.4 * 1000 ng/ml

206400/1900000

=0.11 ng per cells

So the protein present in each cell of Tetrahymena is 0.11ng

write me back if you need more help.

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