• All
  • Primary Antibodies
  • Conjugated Antibodies for IF
  • Conjugated Antibodies for FC
  • Secondary Antibodies
  • Antibody Labeling KitsNew
  • ELISA Kits
  • IHC Kits
  • Magnetic Cell Separation Kits
  • Cytokines & Growth Factors
  • Neutralizing/activating Antibodies
  • Nanobody-based Reagents
  • Accessory Products and Kits
  • Fusion Proteins
×
×
Host
0
Species Reactivity
0
Species Reactivity
0
Conjugate
0
Conjugate
0
Compatible Species
0
Conjugate
0
  • Featured Product
  • KD/KO Validated

CoraLite®594-conjugated Phospho-AKT (Ser473) Monoclonal antibody

Phospho-AKT (Ser473) Monoclonal Antibody for FC (Intra)

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse

Applications

FC (Intra)

Conjugate

CoraLite®594 Fluorescent Dye

CloneNo.

1C10B8

Cat no : CL594-66444

Print datasheet

Synonyms

AKT pan, AKT 1, AKT (pan), AKT, AKT1

验证数据展示

  • FC experiment of PC-3 using CL594-66444

    FC experiment of PC-3 using CL594-66444

    1X10^6 PC-3 cells untreated (dashed line) or treated with Calyculin A (red) were intracellularly stained with 0.25 ug CoraLite®594 Anti-Human Phospho-AKT (Ser473) (CL594-66444, Clone:1C10B8), or 0.25 ug Control Antibody (blue). Cells were fixed and permeabilized with True-Nuclear Transcription Factor Buffer Set.

经过测试的应用

Positive FC (Intra) detected inCalyculin A treated PC-3 cells
Planning an IHC experiment? We recommend our IHCeasy AKT Ready-To-Use IHC Kit. AKT primary antibody included.
For other applications, we recommend the unconjugated version of this antibody, 66444-1-Ig

推荐稀释比

ApplicationDilution
Flow Cytometry (FC) (INTRA)FC (INTRA) : 0.25 ug per 10^6 cells in a 100 µl suspension
It is recommended that this reagent should be titrated in each testing system to obtain optimal results.
Sample-dependent, Check data in validation data gallery.

产品信息

CL594-66444 targets Phospho-AKT (Ser473) in FC (Intra) applications and shows reactivity with human, mouse samples.

Tested Applications FC (Intra) Application Description
Tested Reactivity human, mouse
Immunogen Peptide 种属同源性预测
Host / Isotype Mouse / IgG1
Class Monoclonal
Type Antibody
Full Name v-akt murine thymoma viral oncogene homolog 1
Synonyms AKT pan, AKT 1, AKT (pan), AKT, AKT1
Observed Molecular Weight 60-62 kDa
GenBank Accession NumberNM_005163
Gene Symbol AKT1
Gene ID (NCBI) 207
RRIDAB_2919993
Conjugate CoraLite®594 Fluorescent Dye
Excitation/Emission Maxima Wavelengths588 nm / 604 nm
Form Liquid
Purification MethodProtein A purification
UNIPROT IDP31749
Storage Buffer PBS with 50% Glycerol, 0.05% Proclin300, 0.5% BSA, pH 7.3.
Storage ConditionsStore at -20°C. Avoid exposure to light. Stable for one year after shipment. Aliquoting is unnecessary for -20oC storage.

背景介绍

1) What is AKT?

The serine/threonine kinase B AKT pathway (also known as the PI3K-Akt pathway) plays a vital role in the regulation of cellular processes, including cell proliferation, survival, and growth - processes that are essential for oncogenesis. Mutation of the regulator proteins PI3K and PTEN causes uncontrolled disruption within the PI3-kinase pathway, leading to the development of human cancers (1,2; see also AKT pathway poster for more details).

2) phospho-AKT and FAQs

A)  What is the best way to normalize phosphorylated proteins analyzed by western blot?
Normalize phospho-AKT and total AKT with your loading control (e.g. Actin, tubulin), then calculate the phospho/total ratio using these normalized values.  
Put more simply:
1. Calculate the ratio of band intensities of a phospho-AKT band: the loading control.
2. Calculate the ratio of band intensities of total AKT: loading control.
3. Divide ratio obtained #1 by #2 to obtain a normalized value for comparison among different conditions. This procedure allows one to distinguish between a change in AKT expression and a change in the ratio of phospho-AKT.
* If you are looking at the differences in a phospho-AKT expression resulting from an experimental condition (e.g., knockdown), you should also show the expression of total AKT to distinguish between a change in AKT expression (transcription/translation level) and a change in the AKT phosphorylation status.
B) What is the observed molecular weight for AKT and phospho-AKT?
Molecular Weight AKT - 56 kDa
Molecular Weight phospho-AKT - 60 kDa (Figure 1)
  

Figure 1. WB: HEK-293 cell lysate was subjected to SDS PAGE followed by western blot with 60203-2-Ig (AKT antibody) and 66444-1-Ig (AKT-phospho-S473 antibody) at a dilution of 1:4000 incubated at room temperature for 1.5 hours.

C) Are there any special WB conditions to optimize staining of a phospho-AKT?
Since this is a phosphorylated protein, 5% BSA is recommended over non-fat milk as a blocking agent.
D) What are good positive and negative controls for a phospho-AKT?
- Positive Control: HEK293 cells
- Negative Control: Treatment with PI3K inhibitors (e.g. wortmannin)
E) What species does this antibody react with?

Our internal testing has confirmed that it reacts with the human and mouse forms of phospho-AKT.Reactivity with the human form is also supported by the literature's citations of this antibody.

References:

1. Perturbations of the AKT signaling pathway in human cancer.
2. Targeting the PI3K-Akt pathway in human cancer: rationale and promise.