Paul.xu at 23:13, 25 April 2011

2011-04-25T23:13:13Z

New page

[[Image:MBP--vp16 digestion-4-12 .jpg]]

MBP:42KDa; VP16:55.6Kda;TEV:27 KDa.

MW ladder: 10, 15, 25 (red), 35, 55, 70(red), 100, 130, 250.

 1. NUSA, undigested, from Ni column; 

2. MBP, undigested, from Ni column; 

3. NUSA (1mg/ml ), 1:50 digested by TEV (1mg/ml), from Ni column;

4. MBP(1 mg/ml), 1:50 digested by TEV (1mg/ml) , from Ni column; 

5. PKA_alpha with pGro7, from Ni column; 

6. PKA_alpah with pG-KJE8, from Ni column; 

7. PKA_apha with pKJE7, from Ni column; 

8. PKA-apha with pTf16, from Ni column;

9. PKA-alpha with pG-Tf2, from Ni column;

 

 

 --[[User:Ruili.su|Ruili.su]] 09:36, 13 April 2011 (EST)

for PKA expression: pGro7 and pTf16 are good.

 

[[Image:MBP&NUSA 4-14.jpg]]

 

1. undigested MBP;

2. digested MBP;

3. undigested NUSA;

4. digested NUSA 

--[[User:Ruili.su|Ruili.su]] 09:38, 15 April 2011 (EST)

.[[Image:VP16 purification 2 4 24 2011.jpg]]

1. VP16, Morning elute 

2. Vp16, Afternoon elute 

3. Vp16, afternoon wash 

4. pKA_alpha in pSpeedET with pTF16 chaperone 

5. pKA_alpha in pSpeedET with pKJE7 chaperone 

6. pKA_alpha in pSpeedET with pG-KJE8 chaperone 

 

[[Image:VP16 purification 4 24 2011.jpg]] 

VP16 gel filtration HiLoad 16/60 column, TBS buffer, fration 30 to 35. 

 

[[Image:Vp16 western blot 4 24 11.jpg]]

VP16 western blot, using anti-His tag mouse monoclonal antibody. 

 

 

 

 

--[[User:Paul.xu|Paul.xu]] 12:38, 24 April 2011 (EST) 

SDS-PAGE and western blot analysis of Ruili purified Vp16 

[[Image:Vp16 purification 4 25 11.jpg]] 

Western blot using mouse anti-MBP monoclonal antibody 

[[Image:Vp16 westernblot-2 4 25 11.jpg]] 

Sample # 

 MW ladder 10, 15, 25 (red), 35, 55, 70(red), 100, 130, 250. 

2: vp16 -1

3: vp16- OD280: 0.045

4: VP16 1.29

5: VP16

6: empty

7: MBP: OD280=0.23

8: Morning elute

9: Vp16 in centricon.

So these VP16 proteins purified by Ruili are not VP16, they are MBP.

--[[User:Paul.xu|Paul.xu]] 18:09, 25 April 2011 (EST)

[[Category:Protein_Purification]]

Vp16-purification - Revision history

Paul.xu at 23:13, 25 April 2011