Human p53 marker
(Catalog # B011 )
Description:
Sumoylation marker (human p53 protein) is included to allow clear detection of the somoylation reaction. This assay has been shown to work with crude extracts. Human Aos1/Uba2 (E I), active (Cat# P018; P006), Human UBC9 (E II), active (Cat# P019), and Human SUMO I, active (Cat# P020) are strongly recommended to be used along with this protein.
Assay Conditions:
Sumoylation assays are carried out in a final volume of 20µl in reaction condition (20 mM Hepes pH 7.5, 5mM MgCl2, 2mM ATP). We would recommend to add fresh 2mM ATP to be sure that sufficient energy is supplied.
p53 Sumoylation Control Protocol:
1. Add 1 µl p53 marker (Cat# B011) for each reaction.
2. In each reaction, add 1µl UBC9 (E2) to substrate first, then 1µl Sumo-1, 2µl 10x reaction buffer (Cat# B006), 1µl Aos1/Uba2 (E1). Finally, add H2O to bring up to 20µl (we would recommend to add fresh 2mM ATP to be sure that sufficient energy is supplied).
3. The best reaction concentration of sumoylation proteins is as following:
E I: 7.5µg/ml
E II: 50µg/ml
SUMO-1: 50µg/ml
The recommended condition for control assay is 37ºC for 30~60 minutes.
4. Use 1000x dilution of supplied G S T antibody in 10 ml (Cat# B012) for western blot. One sumoylated band shall be seen on the gel.
Shipping/Storage:
This protein is shipped with dry ice and should be stored at -20ºC or lower.
Legal consideration: FOR RESEARCH USE ONLY.
Price
|
Cat# |
Item | Spec | Price (USD) | Stock location |
|
Polyclonal Antibodies |
||||
|
AP018 |
Sumo I (full length) |
100µl |
300 |
FOB Taiwan |
|
AP021 |
SUMO-1 (11-101) |
100µl |
300 |
FOB Taiwan |
|
AP025 |
100µl |
300 |
FOB Taiwan |
|
|
Proteins |
||||
|
P006 |
Aos1/Uba2 (Good for 100 reactions) |
4.5µg |
150 |
FOB Taiwan |
|
P018 |
Aos1/Uba2 (Good for 100 reactions) |
15µg |
400 |
FOB Taiwan |
|
P019 |
UBC9 (Good for 100 reactions) |
100µg |
100 |
FOB Taiwan |
|
P020 |
SUMO-1 (Good for 100 reactions) |
100µg |
100 |
FOB Taiwan |
|
Kits |
||||
|
K011 |
Convenient Pack |
300 |
FOB Taiwan |
|
|
K010 |
30 reactions |
500 |
FOB Taiwan |
|