Phospho-AKT (Ser473) Monoklonaler Antikörper
Phospho-AKT (Ser473) Monoklonal Antikörper für FC, IHC, WB, ELISA
Wirt / Isotyp
Maus / IgG1
Getestete Reaktivität
human, Maus, Ratte und mehr (5)
Anwendung
WB, IHC, IF, FC, ELISA
Konjugation
Unkonjugiert
CloneNo.
1C10B8
Kat-Nr. : 66444-1-Ig
Synonyme
Galerie der Validierungsdaten
Geprüfte Anwendungen
| Erfolgreiche Detektion in WB | Mit Calyculin A behandelte PC-3-Zellen, Jurkat-Zellen, mit Calyculin A behandelte HEK-293T-Zellen, Mit Calyculin A behandelte HEK-293-Zellen, mit Calyculin A behandelte HSC-T6-Zellen, mit Calyculin A behandelte Jurkat-Zellen, mit Calyculin A behandelte NIH/3T3-Zellen, mit TPA behandelte Jurkat-Zellen, NIH/3T3-Zellen |
| Erfolgreiche Detektion in IHC | humanes Mammakarzinomgewebe, humanes Kolonkarzinomgewebe, mit Calyculin A behandelte Jurkat-Zellen Hinweis: Antigendemaskierung mit TE-Puffer pH 9,0 empfohlen. (*) Wahlweise kann die Antigendemaskierung auch mit Citratpuffer pH 6,0 erfolgen. |
| Erfolgreiche Detektion in FC | Mit Calyculin A behandelte PC-3-Zellen |
Empfohlene Verdünnung
| Anwendung | Verdünnung |
|---|---|
| Western Blot (WB) | WB : 1:2000-1:10000 |
| Immunhistochemie (IHC) | IHC : 1:100-1:400 |
| Durchflusszytometrie (FC) | FC : 0.50 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 | |
Veröffentlichte Anwendungen
| KD/KO | See 1 publications below |
| WB | See 829 publications below |
| IHC | See 58 publications below |
| IF | See 12 publications below |
| FC | See 1 publications below |
Produktinformation
66444-1-Ig bindet in WB, IHC, IF, FC, ELISA Phospho-AKT (Ser473) und zeigt Reaktivität mit human, Maus, Ratten
| Getestete Reaktivität | human, Maus, Ratte |
| In Publikationen genannte Reaktivität | human, Hausschwein, Huhn, Hund, Kaninchen, Maus, Ratte, Zebrafisch |
| Wirt / Isotyp | Maus / IgG1 |
| Klonalität | Monoklonal |
| Typ | Antikörper |
| Immunogen | Peptid |
| Vollständiger Name | v-akt murine thymoma viral oncogene homolog 1 |
| Beobachtetes Molekulargewicht | 60-62 kDa |
| GenBank-Zugangsnummer | NM_005163 |
| Gene symbol | AKT1 |
| Gene ID (NCBI) | 207 |
| Konjugation | Unkonjugiert |
| Form | Liquid |
| Reinigungsmethode | Protein-A-Reinigung |
| Lagerungspuffer | PBS mit 0.02% Natriumazid und 50% Glycerin pH 7.3. |
| Lagerungsbedingungen | Bei -20°C lagern. Nach dem Versand ein Jahr lang stabil Aliquotieren ist bei -20oC Lagerung nicht notwendig. 20ul Größen enthalten 0,1% BSA. |
Hintergrundinformationen
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.
Protokolle
| Produktspezifische Protokolle | |
|---|---|
| WB protocol for Phospho-AKT (Ser473) antibody 66444-1-Ig | Protokoll herunterladen |
| IHC protocol for Phospho-AKT (Ser473) antibody 66444-1-Ig | Protokoll herunterladen |
| Standard-Protokolle | |
|---|---|
| Klicken Sie hier, um unsere Standardprotokolle anzuzeigen |
Publikationen
| Species | Application | Title |
|---|---|---|
Cell Res Inhibiting Hv1 channel in peripheral sensory neurons attenuates chronic inflammatory pain and opioid side effects. | ||
Nat Commun Genome-wide enhancer-gene regulatory maps link causal variants to target genes underlying human cancer risk | ||
Sci Adv Platelet P-selectin initiates cross-presentation and dendritic cell differentiation in blood monocytes. | ||
Acta Pharm Sin B Celastrol targets adenylyl cyclase-associated protein 1 to reduce macrophages-mediated inflammation and ameliorates high fat diet-induced metabolic syndrome in mice. | ||
J Pineal Res Melatonin and verteporfin synergistically suppress the growth and stemness of head and neck squamous cell carcinoma through the regulation of mitochondrial dynamics |
Rezensionen
The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.
FH András (Verified Customer) (07-31-2023) | The Mk-2206 is an allosteric Akt inhibitor that prevents its recruitment to the membrane and consequently its phosphorylation. The Capivasertib is an Akt competitive inhibitor which induces its over phosphorylation.
 |
FH Jorge (Verified Customer) (07-26-2022) | Good signal. Unspecific band below 100 kDa. Used PageRuler Plus Prestained Protein Ladder and chemiluminescence was detected in the 70 kDa marker.
|
FH Tom (Verified Customer) (12-15-2020) | 10ug total protein of HEK293T lysate loaded. Membrane blocked in 5% BSA. Antibody (1:1,000) incubated overnight in block at 4 degrees. Anti-mouse HRP used at 1 in 10,000 to detect band.
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