Description
| Source |
Applications |
Cross-reactivity |
Antibody Type |
M.W. of Akt |
| Rabbit |
WB, IP, IF |
H, M, R, C, Hm, Dro |
Akt C-terminal |
~60kD |
WB, Western blot; IP, Immunoprecipitation; IF, Immunofluorescence; F, Flow cytometry.
H, human; M, mouse; R, rat; C, chicken; Hm, hamster; Dro, drosophila.
This product is a high-purity polyclonal antibody obtained by immunizing rabbit with a synthetic mouse Akt C-terminal segment with appropriate modifications and then purified in two steps using protein A and antigen peptide affinity chromatography.
This Akt antibody recognizes all Akt, detects endogenous Akt1, Akt2, and Akt3, and shows no cross-reactivity with other kinases.
Akt, also known as PKB or Rac, plays an important role in cell survival and apoptosis. Growth and survival factors such as insulin can activate the Akt signaling pathway. Ser473 of Akt can be phosphorylated by PDK1. The PI3 Kinase-Akt signaling pathway is a classical signaling pathway, and inhibition of PI3 kinase by inhibitors, such as LY294002, usually inhibits Akt activation.
A Primary Antibody Dilution Buffer is provided with this product and can be used for primary antibody dilution in Western blot or other applications.
The recommended dilution factors of this product in different applications are as follows. They can be adjusted appropriately according to the level of antigen in samples.
| WB |
IP |
IF |
F |
| 1:1000 |
1:50 |
1:200 |
1:50 |
When using for routine Western blot, this product is sufficient for at least 10 blots.
Packing List
| Item |
Component |
Quantity |
| AA326-1 |
Akt Rabbit Polyclonal Antibody |
10μl |
| AA326-2 |
Primary Antibody Dilution Buffer for Western Blot |
1ml |
| Manual |
– |
1 copy |
Storage Conditions：
Store the Antibody at -20℃ and Primary Antibody Dilution Buffer at 4℃ for up to one year. The Primary Antibody Dilution Buffer can be stored at -20℃ for lone-term storage, but freeze-thaw may result in slight turbidity and a small amount of insoluble substance.
Precautions：
For Western blot analysis, membranes should be incubated overnight in the diluted primary antibody at 4℃ with shaking slowly. Shorter incubation will be less effective.
This product can be recovered for multiple reuses (at least 10 times) in Western blot (WB). Store the unused or recovered diluted antibodies at 4℃.
The recovered antibodies can be reused in the same way as freshly diluted antibodies. If slight turbidity appears, centrifuge the antibody at 10,000×g for 1-3 minutes and use the supernatant for subsequent assays. But it should be discarded when there is significant flocculation or any microbial growth.
This product is for R&D only. Not for drug, household, or other uses.
For your safety and health, please wear a lab coat and disposable gloves during the operation.
Instruction
Instructions for Use：
1. Western blot
a. Dilute an appropriate amount of the Antibody with the provided Primary Antibody Dilution Buffer to an appropriate concentration as required.
b. Transfer the blocked membrane into the diluted primary antibody and incubate at room temperature for 2 hours or overnight at 4℃ with gently shaking. The membrane should be covered completely by the diluted antibody.
c. Recover the diluted primary antibody and store at 4℃ for later use.
d. Proceed to next step in Western blot. For detailed procedures of Western blot, please refer to http://www.beyotime.com/support/western.htm.
2. Immunostaining
For Immunostaining, we recommend using the Immunol Staining Primary Antibody Dilution Buffer (Beyotime, P0103) to dilute the Antibody. The diluted antibody can be recovered for multiple reuses. For detailed procedures of Immunostaining, please refer to http://www.beyotime.com/support/immunol-staining.htm.
3. For other applications, please refer to appropriate protocols.
References
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