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Phospho-Akt-S473 Rabbit mAb (AP1453)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Mouse, Rat

Western blot analysis of lysates from Jurkat, NIH/3T3, C6 cells using Phospho-Akt-S473 Rabbit mAb (AP1453) at 1:1000 dilution. Jurkat, NIH/3T3 and C6 cells were treated by Calyculin A (100 nM) at 37℃ for 30 minutes.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25 μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 20s.

ABclonal:Immunohistochemistry - Phospho-Akt-S473 Rabbit mAb (AP1453)

Immunohistochemistry analysis of paraffin-embedded Mouse brain tissue using Phospho-Akt-S473 Rabbit mAb (AP1453) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse colon tissue using Phospho-Akt-S473 Rabbit mAb (AP1453) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse testis tissue using Phospho-Akt-S473 Rabbit mAb (AP1453) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat kidney tissue using Phospho-Akt-S473 Rabbit mAb (AP1453) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat intestine tissue, Untreated (left) and lambda phosphatase-treated (right), using Phospho-Akt-S473 Rabbit mAb (AP1453) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse spleen tissue, Untreated (left) and lambda phosphatase-treated (right), using Phospho-Akt-S473 Rabbit mAb (AP1453) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Overview

Product name	Phospho-Akt-S473 Rabbit mAb
Catalog No.	AP1453
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC5023-02

Background

The serine-threonine protein kinase encoded by the AKT1 gene is catalytically inactive in serum-starved primary and immortalized fibroblasts. AKT1 and the related AKT2 are activated by platelet-derived growth factor. The activation is rapid and specific, and it is abrogated by mutations in the pleckstrin homology domain of AKT1. It was shown that the activation occurs through phosphatidylinositol 3-kinase. In the developing nervous system AKT is a critical mediator of growth factor-induced neuronal survival. Survival factors can suppress apoptosis in a transcription-independent manner by activating the serine/threonine kinase AKT1, which then phosphorylates and inactivates components of the apoptotic machinery. Mutations in this gene have been associated with the Proteus syndrome. Multiple alternatively spliced transcript variants have been found for this gene. [provided by RefSeq, Jul 2011]

Immunogen information

Immunogen	A synthetic phosphorylated peptide around S473 of human AKT1 (NP_005154.2).
Sequence	HFPQFSYSAS
Gene ID
Swiss Prot
Synonyms	AKT1/AKT2/AKT3; Phospho-Akt-S473
Calculated MW	48kDa/55kDa/51kDa/54kDa
Observed MW	60kDa/

Applications

Reactivity	Human, Mouse, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:500 - 1:1000 IHC-P 1:50 - 1:200 ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements.
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.09% sodium azide, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry
Positive samples	Jurkat treated by Calyculin A, NIH/3T3 treated by Calyculin A, C6 treated by Calyculin A
Cellular location	ciliary basal body, cytoplasm, cytosol, microtubule cytoskeleton, mitochondrion, nucleoplasm, nucleus, plasma membrane, spindle.