Essay on protein laboratory

 DAY: 11/10/12

PROGRAM CODE: BIOL 2365 Relative Biochemistry

SUBJECT: Proteins and Amino Acids

EFFECTS:

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

Amount of Standard Protein/ Unknown (mL)

Absorbance at 750 nm

0

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

Color

X

Bright yellow

Y

Dark yellow

Z

Light magenta

Table several: The outcomes of Test 3; Protein Precipitation

Precipitant

Declaration

TCA

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.

HCl

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

Eluate

Component

Volume collected in cubic centimeters

First

Blue dextrin

three or more. 00

Second

Brown cytochrome

7. 00

Third

Clear phosphate buffer

five. 00

Table 5: The effect of Electrophoresis of Amino Acids

Range moved (+/-) cm Alanine

10% Lactic acid

Phosphate

NH4OH

Glycine

-6. 0

-3. 2/+3. 0

+9. 0

Lysine

-7. 0

-2. 0/0

+3. 7

Glutamic acid

-4. 5

-3. 2/+3. 0

+5. zero

SAMPLE MEASUREMENTS:

(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%)

DISCUSSION:

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. http://amrita.vlab.co.in/?sub=3&brch=63&sim=1094&cnt=1 (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. http://www.phscale.net/pka-ka.htm (13 Oct 2012).

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



News

 Lab Experiment To Study the Frequency Response of 1st Order Low Pass Filtration system Essay

Lab Experiment To Study the Frequency Response of 1st Order Low Pass Filtration system Essay

LAB ASSIGNMENT # 02 NAME: FIRST ORDER HIGH MOVE FILTER. OBJECTIVE: To examine the consistency response of first buy low complete filter.…...

 The Reddish Convertible Composition

The Reddish Convertible Composition

The Reddish colored Convertible In " The Red Convertible”, written by Louise Erdrich, Henry Lamartine makes three unforgettable trips from the Chippewa booking. The initially journey, considered with…...

 Conditional Possibility Essay

Conditional Possibility Essay

Conditional Possibility How to handle Dependent Events Life is full of unique events! You need to get a " feel" so they can be a wise and successful person.…...

 Pt. Multiple Bintang Philippines Research Daily news

Pt. Multiple Bintang Philippines Research Daily news

NGAS Standard Features and Policies Wednesday, March 6, 13 Accrual Accounting • A modified accrual foundation accounting shall be used. •…...

 Essay upon Technology’s Influence on Families

Essay upon Technology’s Influence on Families

Technology's Effect on Families ITS 1000, Laptop or home pc Literacy October 30, 2011 South College or university Online Technology provides a huge effect on families.…...

 Short History Reaction Newspaper

Short History Reaction Newspaper

Brief Story Response Paper Angie Williams ENG/125 March 21 years old, 2011 Shannon Loerch Brief Story Response Paper Everyday Work with, a short history…...