What Is Milliliters Of 20, 000 X A Supernatant?

Submitted By cpandrews
Words: 629
Pages: 3
Open Document
Experiment 4

Post-Lab

Milliliters of 20,000 x g supernatant: 32.5
Grams (NH4)2SO4 used for 40% cut: 7.475
Milliliters of 40% supernatant: 34
Grams of (NH4)2SO4 used for 65% cut: 5.644
Milliliters of 65% supernatant: 35
Milliliters of 65% pellet after resuspending: 3

Fraction Isolated
Sample Assayed μL
Dilution Made
Absorbance Change/min
20,000 x g supernatant
1000
none
.846
(.973-.127)
40% (NH4)2SO4 supernatant
1000
none
.355
(.499-.144)
65% (NH4)2SO4 pellet
1000
none
.662
(1.224-.562)
65% (NH4)2SO4 supernatant
1000
none
.181
(.296-.115)

Bradford Assay

LDH Fraction
Quantity Assayed μL
Dilution Used
Absorbance
20,000 x g supernatant
10
1:20
.352
40% (NH4)2SO4 supernatant
10
1:20
.104
65% (NH4)2SO4 pellet
10
1:20
.275
65% (NH4)2SO4 supernatant
10
1:20
.081

1. Calculation of activity and reaction activity.
Activity= change in absorbance/change minutes x 10^6μM/M x .001L= μmole/min 6200M-1cm-1
For example the 20,000 x g supernatant: .846A/Min x 10^6 μM/M x .001L =.136μmole/min 6200M-1cm-1
The same calculation was carried out for each absorbance; the results are in the table.

Relative Activity=units/volume assayed
For example the 20,000 x g supernatant: .136/1mL=.136

The same calculation was carried out for each absorbance: the results are in the table.
2. Total Units
Total Units= relative activity x total volume fraction
For the first absorbance .136/mL x 32.5mL=4.42 total units
The same calculation was done for each absorbance using the total volume for the fraction. The results are in the table.

3. Protein Concentration
Found using the standard Bradford curve from the previous lab. The chosen protein concentrations are in the following table.

4. Specific Activity
Relative Activity/quantity protein
Ex: for absorbance number 1 .136/16=.0085
The same concept was used for each calculation, in the table below.

Fraction
Units
Units/mL
Total Units
Protein (mg/mL)
Specific Activity
20,000 x g supernatant
.136
.136
4.42
16
.0085
40% (NH4)2SO4 supernatant
.057
.057
1.94
5.9
.0097
65% (NH4)2SO4 pellet .106
.106
.318
13
.0082
65% (NH4)2SO4 supernatant
.029
.029
1.02
5
.0058

Questions
1. The change in absorbance/change in minute was .622. The relative activity would then be .30/3mL times 1/6 which equals .0167.
4. A percent saturated solution of ammonium sulfate is the max amount of solute that can be
Show More

Related Documents: What Is Milliliters Of 20, 000 X A Supernatant?

Example Lab Report Exp 4 Essay

Melissa Chemi 1105 David Pinkston September 28, 2012 Experiment 4: Physical and Chemical Properties PURPOSE: Observe physical and chemical properties of pure substances. PROCEDURE: 1. Place small amounts of a material into 4 test tubes. 2. Observe and record the color and scent of the material. 3. Take one of the test tubes and hold it over a flame using a clamp. Record any reaction the material has to heat. 4. Take a second test tube and fill it ¼ of the way with cold distilled water. Record how…

Words 690 - Pages 3

Exp 1 Lab 0000 4 Essay

Name: Moyo Omoloja Date:2/15/2015 Exp 1: Observations of Chemical Changes Lab Section: 73426 Data Tables: Part 1: Chemicals Well No. Observations of the Reaction A. NaHCO3 + HCl A1 Bubble formation but no color change present B. HCl + BTB B1 Turned a light orange color C. NH3 (NH4OH)+ BTB C1 Turned blue D. HCl + blue dye D1 Turned green E. Blue dye + NaOCl E1 No change occurred after adding NaOCl. with the 1 drop of HCl After adding the HCl a color change occurred. It became 3 different…

Words 754 - Pages 4

Essay about Lecture 3

extremes z z Aslanidis (URV & UNSW) ! ∞, G (z ) ! 0 ! +∞, G (z ) ! 1 Binary Model: Presentation 2 / 14 Logit Models Consider G (xi β) = Λ(xi β) = exp(xi β) 1 + exp(xi β) where G (z ) is the cumulative distribution function for a standard logistic random variable, z. Logit model yi yi Aslanidis (URV & UNSW) = G (xi β) + i exp(xi β) = + 1 + exp(xi β) Binary Model: Presentation i 3 / 14 Probit Models Consider G (xi β) = Φ(xi β) = Z xi β ∞ φ(v )dv where Φ(z ) is the cumulative distribution…

Words 774 - Pages 4

Hw2 Sol Essay

classify x = [0.3, ∗]t . Answer: i )P (ωi ) (a) p(ωi |x) = p(x|ωp(x) . Since P (ω1 ) = P (ω2 ) = P (ω3 ) = 1/3, we just need to compare the likelihood p(x|ωi ). 1 1 exp − [0.3, 0.3][0.3, 0.3]t = 0.1455 2π 2 1 1 p(x|ω2 ) = exp − [−0.7, −0.7][−0.7, −0.7]t = 0.0975 2π 2 p(x|ω1 ) = 2 1 1 exp − [−0.2, −0.2][−0.2, −0.2]t 4π 2 1 1 + exp − [0.8, −0.2][0.8, −0.2]t 4π 2 = 0.1331 p(x|ω3 ) = Since p(x|ω1 ) is the largest likelihood, x…

Words 648 - Pages 3

A Description Of Intern Survey Assistants

Home Survey Assistant-Internship exp - Calgary, AB Internship With a mission to understand, innovate, partner and deliver, exp provides professional, technical and strategic advisory services related to the world's built and natural environments in six key practice areas: Buildings, Earth & Environment, Energy, Industrial, Infrastructure, and Sustainability. Our heritage dates back to 1928, and today more than 3,400 creative professionals across North America and the globe provide the expertise…

Words 428 - Pages 2

WeekfourReflection Team C Essay

Adjustments Adjusted Trial Balance Income Statement Balance Sheet Dr. Cr. Dr. Cr. Dr. Cr. Dr. Cr. Dr. Cr. 1 Cash 37080 37080 37080 2 Accounts receivable 52320 i-600 51720 51720 3 Notes receivable 24000 24000 24000 4 Merchandise inv. 1.105 31650 31650 31650 5 Prepaid insurance 4800 c-2500 2300 2300 6 Prepaid rent 4395 b-2800 1595 1595 7 Store supplies 1650 f-1000 650 650 8 Furniture & fixture 13500 13500…

Words 940 - Pages 4

Midterm Study Guide Essay

A/R(Net value), Inventory (LCM), PP Exp (Net book value)Principles of financial accounting: Objectivity: financial acct is verifiable & reliable; Matching: efforts (Exp) should be matched with acc. (Rev) ex. Direct association exists, exp cost when rev. is recognized. If assoc. diff. to identify, allocate costs systematically; no association, expense immediately. Revenue Recognition: rev. recogn when earnings process is virtually comp & transaction occurred, 4 criteria to meet 1) complete sign portion…

Words 926 - Pages 4

Questions On Production In Chile

Families/Kids/Spanish/Argentina/adapted Older/Healthy/Spanish/Argentina/1 yrs old Families/Economy/Spanish/Argentina/1 yrs old Families/Kids/Spanish/Argentina/1 yrs old Channel Total Exp. (mill.) Traditional Self Serve Hypermarket Web/Other Home Wholesale Distribute? Yes Yes Yes Yes Yes Allow. 7.6% 7.6% 7.6% 7.6% 7.6% 7.6% 7.6% 7.6% 7.6% Sales People ARS8.3 5 3 9 3 4 Sales Forecast 2.1 1.8 3.7 2.1 4.3 7.0 3.5 3.0 2.0 Budget (mill.) 5.0 7.0 2.0 not in use not in use not in use Promote? ARS3.8 Yes Yes Yes No No Allocate…

Words 741 - Pages 3

The Deserted Village Essay

would waste these things(health ,happiness and power). The best way to keep the flame of his life from wasting is to get back to his birth place (village). 17th and 18th line _attend→ feel _pride→ part and parcel of human life _swains→ villagers Exp: The poet now is away from the village , but he has a dream (wish). Human beings feel proud of themselves sometimes, this is human quality, so he says that as long as he is a human being he wants to show his pride in his knowledge . When the poet left…

Words 2027 - Pages 9

Case 3 Lilian Ltd Essay examples

Executive Summary Lilian Lagat is a woman who set up her real estate and property management business in 2007. In six years she became successful and reliable business person. Now she wants to make a right decision to invest her money into hotel business in Eldoret town, Kenya. Earlier Eldoret was a capital of Kenya and now is the largest town in western Kenya. It is on the 15th place among large cities in the whole country. The population is pretty high and there is Moi University in Eldoret with…

Words 1891 - Pages 8