Clone:
REA359
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
ICFC, MC
Alternative names:
Akt1, CWS6, PKB, PKB-ALPHA, RAC, RAC-ALPHA, PKBγ, PKBβ, STK-2

Extended validation for AKT pS473 Antibody, anti-human/mouse, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA359
M89-61++
Cells were incubated with an excess of purified unconjugated AKT pS473 (REA359) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
Flow cytometric comparison of different clones for AKT pS473. Jurkat cells were first stained with Viobility Fixable Dye followed by fixation and permeabilization using the Cell Signaling Buffer Set A . Cells were the stained with AKT pS473 antibodies and plotted against the side scatter. As a control, antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for AKT pS473. Jurkat cells were first stained with Viobility Fixable Dye followed by fixation and permeabilization using the Cell Signaling Buffer Set A . Cells were the stained with AKT pS473 antibodies and plotted against the side scatter. As a control, antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for AKT pS473. Jurkat cells were first stained with Viobility Fixable Dye followed by fixation and permeabilization using the Cell Signaling Buffer Set A . Cells were the stained with AKT pS473 antibodies and plotted against the side scatter. As a control, antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for AKT pS473. Jurkat cells were first stained with Viobility Fixable Dye followed by fixation and permeabilization using the Cell Signaling Buffer Set A . Cells were the stained with AKT pS473 antibodies and plotted against the side scatter. As a control, antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals and Viobility 405/520 Fixable Dye fluorescence. The recommended titers of respective antibodies from different suppliers were used.

Specifications for AKT pS473 Antibody, anti-human/mouse, REAfinity™

Overview

Clone REA359 recognizes the human and mouse Akt antigen phosphorylated at serine 473 (pS473). Akt is a family of serine/threonine kinases which is also known as protein kinase B (PKB) or related to the A and C kinases (RAC-PK). Akt intricately regulates many cellular functions such as cell growth and proliferation, cell survival, apoptosis, energy metabolism, and resistance to anticancer therapeutics. It consists of three isoforms, Akt1, Akt2, and Akt3 (or PKBα, β, and γ). Although these three isoforms are encoded by separate genes, they share a common NH2-terminal pleckstrin homology (PH) domain, a catalytic domain in the middle, and a COOH terminus. The identity of the overall amino acid sequence of the three isoforms is very high (about 80%); however, the COOH terminus and the PH-linker region are more diverse. All three isoforms contain similar phosphorylation/activation sites: threonine 308 (Akt1), 309 (Akt2), and 305 (Akt3) and serine 473 (Akt1), 474 (Akt2), and 472 (Akt3). Phosphorylated or activated Akt regulates many of its cellular functions through the phosphorylation of a series of downstream players.
Additional information: Clone REA359 displays negligible binding to Fc receptors.

Alternative names

Akt1, CWS6, PKB, PKB-ALPHA, RAC, RAC-ALPHA, PKBγ, PKBβ, STK-2

Detailed product information

Technical specifications

CloneREA359
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (I), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, mouse
AntigenAKT pS473
Alternative names of antigenAkt1, CWS6, PKB, PKB-ALPHA, RAC, RAC-ALPHA, PKBγ, PKBβ, STK-2
Molecular mass of antigen [kDa]56
Distribution of antigenother
Entrez Gene ID11651, 207
RRIDAB_2751826, AB_2751791, AB_2751731, AB_2801919, AB_2651213, AB_2651214, AB_2651215, AB_2651216, AB_2751857

References for AKT pS473 Antibody, anti-human/mouse, REAfinity™

Publications

  1. Calamito, M. et al. (2010) Akt1 and Akt2 promote peripheral B-cell maturation and survival. Blood 115: 4043-4050
  2. Arranz, A. et al. (2012) Akt1 and Akt2 protein kinases differentially contribute to macrophage polarization. Proc. Natl. Acad. Sci. U.S.A. 109(24): 9517-9522
  3. Santi, S. A. et al. (2010) The Akt isoforms are present at distinct sub cellular locations. Am. J. Physiol., Cell Physiol. 298(3): 580-591

Related products for
AKT pS473 Antibody, anti-human/mouse, REAfinity™

3 products available