2013 Promega catalogue

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2013
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Cell Signaling
Anti-pS 473 Akt pAb
Product Size Cat.# Price ($)
Anti-pS 473 Akt pAb 40 μl G7441 505
For Research Use Only. Not for Use in Diagnostic Procedures.
Description: Anti-pS 473 Akt pAb is an affinity-purified polyclonal rabbit
antibody. The antibody is purified using a phosphorylated peptide that
corresponds to the phospho-S 473 form of Akt-1 and is useful for both Western
blotting and immunocytochemistry.
Features:
• Specificity: The antibody is selective for the Ser 473 phosphorylated
isoforms of Akt and does not show cross-reactivity with nonphosphorylated
Akt.
• Immunogen: Peptide from the singly phosphorylated Ser 473 region from
the C-terminus of Akt-1 protein.
• Antibody Form: Affinity-purified rabbit IgG, supplied in PBS with 50μg/ml
gentamicin.
• Value: Will generate 100ml of blotting solution, sufficient for 10 Western
blots of 10ml each.
Storage Conditions: Store at 4°C for daily/weekly use or dispense into
aliquots and store at –20°C for long-term storage.
Protocol
Anti-pS 473 Akt pAb Protocol
Part#
9PIG744
A. 1 2
B.
1 2
– Phospho-Akt – Akt
Detection of phosphorylated Akt by Western blot analysis with
Anti-pS 473 Akt pAb. Panel A. NIH/3T3 total cell extract (10µg per lane)
was resolved by polyacrylamide gel electrophoresis and blotted onto
nitrocellulose. Lane 1, untreated cells; lane 2, cells pretreated with PDGF
(Invitrogen) at 50ng/ml for 20 minutes. Anti-pS 473 Akt pAb (Cat.# G7441)
was used at a 1:2,500 dilution. The blot was probed with Donkey
Anti-Rabbit IgG (H+L), HRP, Anti-ACTIVE ® Qualified pAb (Cat.# V7951) at
1:10,000 dilution followed by chemiluminescent detection. Panel B. A
pan-Akt pAb (New England Biolabs) reveals total Akt in both stimulated
and unstimulated NIH/3T3 cell extracts. Secondary antibody and detection
methods were the same as those used in Panel A.
2810TA10_9A
2769TA10_9A
Immunocytochemical staining of Akt in embryonic rat brain cells.
Embryonic (day 17) rat brain cells were collected and treated with
20ng/ml each of EGF and FGF. Anti-pS 473 Akt pAb was used at a 1:50
dilution. Positive cells were visualized using a donkey anti-rabbit, Cy ® 3-
conjugated secondary antibody. Nuclei were stained using DAPI. Protocols
developed and performed at Promega.
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