Lab: Western Blot DOX DO DOXDOX Cont 1 40 2 303 20r 50 BCL SDHA -AC TIN MT-COx 1
ID: 210014 • Letter: L
Question
Lab: Western Blot DOX DO DOXDOX Cont 1 40 2 303 20r 50 BCL SDHA -AC TIN MT-COx 1 37 16 The above image is a hypothetical western blot from an experiment testing the cancer killing ability of the anti-cancer drug doxorubicin (DOX) at various concentrations ranging from 10-40uM. Three protein bands appear on the blot (BCL-2, BAX, and -actin). The protein BCL-2 is anti-apoptosis and typically promotes cell survival, which is conducive for cancer development. BAX on the other hand promotes apoptosis and induces increased cell death. Answer the following questions completely and to the best of your ability and include with your respective lab report. 1. Based upon the results, what can one infer about the anti cancer effectiveness of the chemotherapeutic drug DOx? Are increasingly higher doses of the anti-cancer drug more or less effective? 2. Approximate the size of each of the protein bands in the above western blot image 3. Explain how you would use the protein standard/ladder in combination with the chosen primary and secondary antibodies to identify the protein bands.Explanation / Answer
1. Observation: In the above results Bax (promotes apoptosis and induces cell death) is not expressed in the cancer cells when the drug dosage is lower (10 micro M). Its expression level gradually increases with increase in drug dosage through 20 and 30 microM and it is highly expressed at maximum drug dose concentration of 40 micro M.
The expression level of Bcl-2 (anti-apoptotic, promoting cancer cell survial) remains same at any concentration of doxorubicin, But at 40 micro M, the expression of Bax looks stronger than Bcl-2
Presence of Bax induces death of cancer cells and helps in prevention of cancer cell growth.
Inference: The drug doxorubicin is more effective with increased doses where it increases the apoptotic cell death of cancer cells.
2. according to given picture,
Appx size of beta-actin band = 42kDa
Appx size of Bax with its marker MT-COX1= 40kDa
3. Protein standard/ladder can be used to identify protein bands by knowing the molecular weights of chosen primary or secondary antibodies. The protein standard will have the marker bands with known protein molecular weights. This can be compared with the protein of our interest and identify it.
Appx size of Bcl-2 with its marker SDHA = 67 kDa
Related Questions
Navigate
Integrity-first tutoring: explanations and feedback only — we do not complete graded work. Learn more.