Essay on protein laboratory

 DAY: 11/10/12

PROGRAM CODE: BIOL 2365 Relative Biochemistry

SUBJECT: Proteins and Amino Acids


Table 1: The results of experiment 1; the Lowry Test

Amount of Standard Protein/ Unknown (mL)

Absorbance at 750 nm


0. 000

zero. 1

zero. 017

0. 3

zero. 135

0. 3

zero. 155

0. 5

0. 230

zero. 7

zero. 323

zero. 7

zero. 310

1 . 0

0. 457

1 ) 0 Unfamiliar 1a

0. 463

1 . 0 Unfamiliar 1b

0. 433

1 ) 0 Unknown 2a

zero. 237

1 . 0 Unknown 2b

zero. 159

Table 2: The results of Experiment 2; Ninhydrin Test out

Amino acid



Bright yellow


Dark yellow


Light magenta

Table several: The outcomes of Test 3; Protein Precipitation




White comfortable precipitate in large mass, the comfortable mass goes from the bottom towards the top of the conduit. The solution was thin and loose. Ammonium sulphate

White gel-like medications, the solution was thick.


White precipitate accumulated towards the top and also the underlying part of the solution. The solution was thin. NaOH

No medications seen, the perfect solution is was cream in color.

Copper sulphate

Light blue coagulated precipitate viewed at the top of the solution. Lead Acetate

Cloudy remedy with little specks of white medicine.

Ice cold ethanol

Cloudy at the end and about zero. 5 cm of the the top of solution was clear.

Desk 4: The results of Gel Purification Chromatography



Volume collected in cubic centimeters


Blue dextrin

three or more. 00


Brown cytochrome

7. 00


Clear phosphate buffer

five. 00

Table 5: The effect of Electrophoresis of Amino Acids

Range moved (+/-) cm Alanine

10% Lactic acid




-6. 0

-3. 2/+3. 0

+9. 0


-7. 0

-2. 0/0

+3. 7

Glutamic acid

-4. 5

-3. 2/+3. 0

+5. zero


(1) Necessary protein concentration of Standard Protein Solution:

1mL = 200µg protein

Consequently 0. 1mL= 0. 1/1× 200 = 20µg

(2) Protein attentiveness of the two unknowns:

Gradient of the range = Y2-Y1/ X2-X1

zero. 135-0. 017/ 60-20 = 0. 00295

Y= mx

Average absorbance of Unfamiliar 1= zero. 463 + 0. 433/ 2 sama dengan 0. 448

Therefore: zero. 448 = 0. 00295 x

x = zero. 448/ zero. 00295 = 151. 86µg

Therefore 1mL of Unidentified 1 provides 151. 86µg of necessary protein

Average absorbance of Unknown 2 sama dengan 0. 237 + zero. 159/ 2 = zero. 198

Consequently: 0. 198 = zero. 00295 x

x sama dengan 0. 198/ 0. 00295 = 67. 12µg

Consequently 1mL of Unknown a couple of has 67. 12µg of protein

(3) The void volume is the volume among and outside the gel beans in the steering column. This quantity is therefore calculated making use of the eluate value of the largest molecule (macromolecule) that was completely ruled out from the steering column. This is thus since the most significant molecule takes up the areas between and outside the skin gels beads being that they are too large to and thus that they travel out more quickly than the smaller molecules which can become trapped in the pores with the beads. Which means total amount of this particle will reveal the void volume. The macromolecule in this experiment was blue dextran since it was eluated first. The amount of green dextran was 3. 00mL; therefore the gap volume was also several. 00mL. (4) Bed volume = amount of a canister = π r2h

Where h = 15cm r = 0. 5cm and π sama dengan 3. 18

Bed amount = 14. 78 cm3

(5) The fraction of your bed volume that is void volume level = several. 00/ 14. 78 = 0. 254 (25. 4%)


The Lowry Assay is known as a biochemical check that assists in the perseverance of the total amount of protein present in a solution. The principle of the assay is grounded around the reactivity with the peptide nitrogen with copper mineral ions in alkaline circumstances followed by a discount of the Folin- Ciocalteu's phosphomolybdiphosphotungstic acid; which can be yellow in color to heteropolymolybdenum which can be blue in color. This can be done by way of an oxidation of fragrant protein residues by copper (Boyer 2012). The reaction is really as follows:

Figure 1: the Lowry effect

A spectrophotometer was used to measure the color change from the Folin- Ciocalteu's reagent; this absorbance worth is then valuable in determining...

Referrals: Amrita College or university. Qualitative Analysis of Proteins. 2012. (4 August 2012)

Boyer, Rodney. 2012. Biochemistry Lab. Pentice Lounge Publishers, Nj-new jersey.

Nelson, Cox. 2008. Leningher Principles of Biochemistry. 6th edition. W. H Freeman and Firm, New York.

Nigam Aarti, Ayyahguri. 2012. Archana Lab Manual in Biochemistry, Immunology and Biotechnology. Acara susunan acara Mc Graw Hill Creating Company.

pHscale. Net, pKa and Ka, 2012. (13 Oct 2012).

University of Cal, Department of Chemistry. 2011. Chemistry 2B Laboratory Manual. University of California. Davis, CA.


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