Homework Answers
16.
a. Lane 4 contains the protein of interest.
b. The protein is insoluble because it has been detected in lane 4 which is insoluble fraction.
c. The protein visible on western blot is around 20kDa. So the target protein is the protein which is showing molecular weight around 20kDa.
d. Molecular weight of protein of interest is around 20kDa.
e. I can see there are two contaminants in the fraction from lane 4. The upper contaminant is around 36-38kDa and lower is around 10 kDa.
f. We have to keep elution since it contains protein of interest.
g. Cation exchange column is used to purify positively charged protein.
So, charge on protein in Lane 5- negative and in Lane 6- positive
Add Answer to:
Betergents h. What chemical would you add to refold this protein? 16. You wish to purify...
Proteins: BSA, yeast ADH, ovalbumin, lysozyme . Discuss how you were able to determine which protein...
Proteins: BSA, yeast ADH, ovalbumin, lysozyme
. Discuss how you were able to determine which
protein was in which lane. You will have to do some
online research to find the molecular weight of each protein,
whether it is a dimer, etc. For at least one of the proteins, there
is some variability regarding the molecular weight, but the range
should still allow you to determine which lane each of the proteins
is in. For each lane, pay attention to...
0.5. What fraction of the offsprings of two parents with Sickle cell trait would you expect...
0.5. What fraction of the offsprings of two parents with Sickle cell trait would you expect to have Sickle cell anemia? Q.6. List the major steps you undertake for performing ELISA? 0.7. Describe an ELISA procedure you'll undertake to determine if a person is infected with AIDS virus? Q.3. In the two protein gel electrophoresis labs done in this class, proteins were separated based on their net charge or their size (weight). Explain similarities and differences between these two laboratory...
You have a solution containing human myoglobin (16.7 KD, pi7.0), hemoglobin (64.5 KD; pi7.1). and serum...
You have a solution containing human myoglobin (16.7 KD, pi7.0), hemoglobin (64.5 KD; pi7.1). and serum albumin (66,5 KD: pl 4,9). You want to separate these proteins and you run one portion of the solution through anion exchange chromatography at pH 8 and a second portion of the solution through gel filtration. You measure the protein concentration in fractions collected from both columns and you get the following elution profiles Anion Exchange filtration Protein Concentration Eurim Volume Elution Volume Based...
Hi can you check/help me with these? And please explain if you can! Thank you! These...
Hi can you check/help me with these? And please explain if you
can! Thank you!
These questions are about Ion-exchange
Chromatography
1. The charge on the column is negative
a. Would the protein KLGVRQYWRRRRPLYWTV bind to it?
2. Does the buffer action work by a difference in ionic
strength?
3.
4. T/F: You will likely get more fractions of DNP-glycine than
the other two proteins.
True because DNP-glycine goes through very fast because
it is small.
5. T/F: Cytochrome c...
Why are proteins heat denatured prior to analysis in SDS-PAGE? Select all answers that apply. Denaturation...
Why are proteins heat denatured prior to analysis in SDS-PAGE? Select all answers that apply. Denaturation of the protein is necessary so that proteins run proportionally to their size, based on the interaction of SDS with the unfolded protein. Heat is used to hydrolyze the peptide bonds of the protein. The heat step allows the proteins to unfold, enabling the protein chain to be coated with SDS molecules. Heat is used to hydrolyze disulfide bonds. QUESTION 2 1.5 points Saved...
Q 4. Explain how Fig.1A (arrow) result corroborates with the author hypothesis about Parkin aggre...
Q
4. Explain how Fig.1A (arrow) result corroborates with the author
hypothesis about Parkin aggresome and proteasome activity in PD?
MG-132+ HMW 127- 84-- 41- DTT+ IP T HMW 6- IP : anti-flag WB: anti-ubiquitin Fia. 1. Parkin forms plexes A, HEK 293Tells transiently tranafected with FLAG-Parkin were divided into twe dishes and were either untreated or treated with 5PM MG-132 for 161). Cells were then lysed abidใหr-taining 1% Triton X-100 ลnd fractitated into "Iuble (S) and insoluble u) frac...
4. (Challenge) You are purifying a protein from the brain of patients with an unusual type of...
4. (Challenge) You are purifying a protein from the brain of patients with an unusual type of dementia triggered by a coronavirus and you hypothesize that this protein may be a signature molecule of this type of dementia. To pursue this hypothesis, you collect brain tissue from patients post mortem, do differential centrifugation, density gradient centrifugation, CM (carboxymethyl cellulose) ion exchange chromatography and gel filtration. You then assess the purity of your protein using both native and SDS gel electrophoresis. The...
please answer all that you can 1. You have genetically engineered green fluorescent protein (GFP) containing...
please answer all that you can 1. You have genetically engineered green fluorescent protein (GFP) containing a KDEL sequence (GFP-KDEL). When GFP-KDEL is expressed in normal human fibroblasts and examined using fluorescence microscopy, the fluorescence appears diffuse across the cytoplasm. How would you explain this observations given that KDEL is supposed to be an ER-specific sorting sequence? A. This engineered GFP would not have a hydrophobic signal sequence to get it into the RER in the first place. B. The...
QUESTION 6 Assume you are studying a protein-coding gene, ACEX, which includes 4 exons as illustrated...
QUESTION 6 Assume you are studying a protein-coding gene, ACEX, which includes 4 exons as illustrated in the gene map below. The 5' UTR and 3' UTR segments are each 25 bp long. Exons 1 thru 4 are 100, 200, 300, 400 bp long, respectively. Each intron is 200 bp each. The locations of the relevant EcoRI sites within the ACEX locus are indicated, but the location of other restriction enzyme sites (like BamHI) are not shown." EcoRI probe EcoRI...
. Gleevec (Imatinib) inhibits protein kinase BCR-ABL, which is constitutively active in patients with Chronic Myelogenous...
. Gleevec (Imatinib) inhibits protein kinase BCR-ABL, which is
constitutively active in patients with Chronic Myelogenous Leukemia
(CML). The structure of Gleevec is shown below:
a) Based on our discussion in lecture and the structure above,
explain how the “lead” compound structure for Gleevec was designed
and how it led to the development a compound that could bind ABL
with higher affinity than ATP.
b) Using a broad kinase inhibition assay, you discover that
Gleevec inhibits another tyrosine kinase called...
Proteins: BSA, yeast ADH, ovalbumin, lysozyme
. Discuss how you were able to determine which
protein was in which lane. You will have to do some
online research to find the molecular weight of each protein,
whether it is a dimer, etc. For at least one of the proteins, there
is some variability regarding the molecular weight, but the range
should still allow you to determine which lane each of the proteins
is in. For each lane, pay attention to...
0.5. What fraction of the offsprings of two parents with Sickle cell trait would you expect to have Sickle cell anemia? Q.6. List the major steps you undertake for performing ELISA? 0.7. Describe an ELISA procedure you'll undertake to determine if a person is infected with AIDS virus? Q.3. In the two protein gel electrophoresis labs done in this class, proteins were separated based on their net charge or their size (weight). Explain similarities and differences between these two laboratory...
You have a solution containing human myoglobin (16.7 KD, pi7.0), hemoglobin (64.5 KD; pi7.1). and serum albumin (66,5 KD: pl 4,9). You want to separate these proteins and you run one portion of the solution through anion exchange chromatography at pH 8 and a second portion of the solution through gel filtration. You measure the protein concentration in fractions collected from both columns and you get the following elution profiles Anion Exchange filtration Protein Concentration Eurim Volume Elution Volume Based...
Hi can you check/help me with these? And please explain if you
can! Thank you!
These questions are about Ion-exchange
Chromatography
1. The charge on the column is negative
a. Would the protein KLGVRQYWRRRRPLYWTV bind to it?
2. Does the buffer action work by a difference in ionic
strength?
3.
4. T/F: You will likely get more fractions of DNP-glycine than
the other two proteins.
True because DNP-glycine goes through very fast because
it is small.
5. T/F: Cytochrome c...
Why are proteins heat denatured prior to analysis in SDS-PAGE? Select all answers that apply. Denaturation of the protein is necessary so that proteins run proportionally to their size, based on the interaction of SDS with the unfolded protein. Heat is used to hydrolyze the peptide bonds of the protein. The heat step allows the proteins to unfold, enabling the protein chain to be coated with SDS molecules. Heat is used to hydrolyze disulfide bonds. QUESTION 2 1.5 points Saved...
Q
4. Explain how Fig.1A (arrow) result corroborates with the author
hypothesis about Parkin aggresome and proteasome activity in PD?
MG-132+ HMW 127- 84-- 41- DTT+ IP T HMW 6- IP : anti-flag WB: anti-ubiquitin Fia. 1. Parkin forms plexes A, HEK 293Tells transiently tranafected with FLAG-Parkin were divided into twe dishes and were either untreated or treated with 5PM MG-132 for 161). Cells were then lysed abidใหr-taining 1% Triton X-100 ลnd fractitated into "Iuble (S) and insoluble u) frac...
QUESTION 6 Assume you are studying a protein-coding gene, ACEX, which includes 4 exons as illustrated in the gene map below. The 5' UTR and 3' UTR segments are each 25 bp long. Exons 1 thru 4 are 100, 200, 300, 400 bp long, respectively. Each intron is 200 bp each. The locations of the relevant EcoRI sites within the ACEX locus are indicated, but the location of other restriction enzyme sites (like BamHI) are not shown." EcoRI probe EcoRI...
. Gleevec (Imatinib) inhibits protein kinase BCR-ABL, which is
constitutively active in patients with Chronic Myelogenous Leukemia
(CML). The structure of Gleevec is shown below:
a) Based on our discussion in lecture and the structure above,
explain how the “lead” compound structure for Gleevec was designed
and how it led to the development a compound that could bind ABL
with higher affinity than ATP.
b) Using a broad kinase inhibition assay, you discover that
Gleevec inhibits another tyrosine kinase called...
Most questions answered within 3 hours.
-
Decision #2: Planning for
Retirement
Luke and Olivia are 22, newly married, and ready to embark...
asked 1 minute ago -
Which one of these statements is correct concerning the time
value of money?
Decreasing the PV...
asked 1 minute ago -
Imagine a PPF for two goods: guns and butter. A natural disaster
hits and reduces our...
asked 2 minutes ago -
Two wavefronts that add with constructive amplitude
reinforcement, have what property?
There are 270° out of...
asked 5 minutes ago -
Find the variance for the given data. Round your answer to one
more decimal place than...
asked 7 minutes ago -
Electromagnetic radiation of wavelength 96.4 nm was projected on
a zinc plate with a 6.01×10-19 J...
asked 23 minutes ago -
5. Explain what happens when the dose is larger than the
threshold dose.
6. Explain how...
asked 15 minutes ago -
I have a few concentrations that are in mg/L and I need to
calculate them in...
asked 17 minutes ago -
My son, daughter and dog have successfully built a storm-making
machine! They've created a storm that...
asked 23 minutes ago -
Q. The neighborhood kids set up an outdoor lemonade stand in
Maryland in June. They find...
asked 36 minutes ago -
Solution, dilution, concentration problems: Please answer 1-5
and show all work
1. How many grams of...
asked 43 minutes ago -
Based on a predicted level of production and sales of 22,000
units, a company anticipates total...
asked 53 minutes ago