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Phospho-AKT (S473) Recombinant Rabbit Monoclonal Antibody [PSH24-43]

Usd: 385

Specification

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Catalog# HA724447

Phospho-AKT (S473) Recombinant Rabbit Monoclonal Antibody [PSH24-43]

Application

  • WB

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

Phospho-AKT (S473) Recombinant Rabbit Monoclonal Antibody [PSH24-43]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic phospho-peptide corresponding to residues surrounding Ser473 of human Akt1.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IP

Molecular Weight

Predicted band size: 56 kDa

Positive Control

293T (Human embryonic kidney cells) cell lysate, 293T treated with 100 nM Calyculin A for 15 minutes cell lysate, C2C12 treated with 100 nM Insulin for 30 minutes cell lysate, C6 (Rat glioma cell) cell lysate, C6 treated with 50 ng/mL Calyculin A for 45 minutes cell lysate.

Conjugation

unconjugated

Clone Number

PSH24-43

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IP

  • 1-2μg/sample

Target

Function

RAC(Rho family)-alpha serine/threonine-protein kinase is an enzyme that in humans is encoded by the AKT1 gene. This enzyme belongs to the AKT subfamily of serine/threonine kinases that contain SH2 (Src homology 2-like) protein domains. It is commonly referred to as PKB, or by both names as "Akt/PKB". The serine-threonine protein kinase AKT1 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.

Background References

1. Lee DS et al. P2 × 7 Receptor Inhibits Astroglial Autophagy via Regulating FAK- and PHLPP1/2-Mediated AKT-S473 Phosphorylation Following Kainic Acid-Induced Seizures. Int J Mol Sci. 2020 Sep

2. Cai Q et al. MAPK6-AKT signaling promotes tumor growth and resistance to mTOR kinase blockade. Sci Adv. 2021 Nov

Subcellular Location

Cytoplasm, Nucleus, Cell membrane.

UNIPROT

SwissProt: P31749 Human

SwissProt: P31751 Human

SwissProt: Q9Y243 Human

SwissProt: P31750 Mouse

SwissProt: Q60823 Mouse

SwissProt: Q9WUA6 Mouse

SwissProt: P47196 Rat

SwissProt: P47197 Rat

SwissProt: Q63484 Rat

Synonyms

AKT1 antibody

AKT1_HUMAN antibody

MGC99656 antibody

PKB antibody

PKB-ALPHA antibody

PRKBA antibody

Protein Kinase B Alpha antibody

Protein kinase B antibody

Proto-oncogene c-Akt antibody

RAC Alpha antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-AKT (S473) on different lysates with Rabbit anti-Phospho-AKT (S473) antibody (<a href="/products/HA724447" style="font-weight: bold;text-decoration: underline;">HA724447</a>) at 1/1,000 dilution. <br /> <br />Lane 1: 293T (Human embryonic kidney cells) cell lysate <br />Lane 2: 293T treated with 100 nM Calyculin A for 15 minutes cell lysate <br />Lane 3: C2C12 (Mouse myoblasts) cell lysate <br />Lane 4: C2C12 treated with 100 nM Insulin for 30 minutes cell lysate <br /> <br />Lysates/proteins at 15 µg/Lane. <br />Exposure time: 20 seconds; ECL: K1801 <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA724447" style="font-weight: bold;text-decoration: underline;">HA724447</a>, 1/1,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 56 kDa <br />Observed band size: 56 kDa

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-AKT (S473) on different lysates with Rabbit anti-Phospho-AKT (S473) antibody (HA724447) at 1/1,000 dilution.

    Lane 1: 293T (Human embryonic kidney cells) cell lysate
    Lane 2: 293T treated with 100 nM Calyculin A for 15 minutes cell lysate
    Lane 3: C2C12 (Mouse myoblasts) cell lysate
    Lane 4: C2C12 treated with 100 nM Insulin for 30 minutes cell lysate

    Lysates/proteins at 15 µg/Lane.
    Exposure time: 20 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724447, 1/1,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 56 kDa
    Observed band size: 56 kDa

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of Phospho-AKT (S473) on different lysates with Rabbit anti-Phospho-AKT (S473) antibody (<a href="/products/HA724447" style="font-weight: bold;text-decoration: underline;">HA724447</a>) at 1/2,000 dilution. <br /> <br />Lane 1: C6 (Rat glioma cell) cell lysate <br />Lane 2: C6 treated with 50 ng/mL Calyculin A for 45 minutes cell lysate <br /> <br />Lysates/proteins at 10 µg/Lane. <br />Exposure time: 17 seconds; ECL: K1801 <br /> <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br />Primary antibody: <a href="/products/HA724447" style="font-weight: bold;text-decoration: underline;">HA724447</a>, 1/2,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 °C <br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Predicted band size: 56 kDa <br />Observed band size: 56 kDa

    ☑ Cell treatment (CT)

    Western blot analysis of Phospho-AKT (S473) on different lysates with Rabbit anti-Phospho-AKT (S473) antibody (HA724447) at 1/2,000 dilution.

    Lane 1: C6 (Rat glioma cell) cell lysate
    Lane 2: C6 treated with 50 ng/mL Calyculin A for 45 minutes cell lysate

    Lysates/proteins at 10 µg/Lane.
    Exposure time: 17 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA724447, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 °C
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 56 kDa
    Observed band size: 56 kDa

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunoprecipitation (IP) <br /> <br />Phospho-AKT (S473) was immunoprecipitated in 0.2 mg NIH/3T3 (Mouse fibroblast) treated with 100 nM Calyculin A for 30 minutes cell lysate with <a href="/products/HA724447" style="font-weight: bold;text-decoration: underline;">HA724447</a> at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using <a href="/products/HA724447" style="font-weight: bold;text-decoration: underline;">HA724447</a> at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. <br /> <br />Lane 1: NIH/3T3 treated with Calyculin A cell lysate (input) <br />Lane 2: <a href="/products/HA724447" style="font-weight: bold;text-decoration: underline;">HA724447</a> IP in NIH/3T3 treated with Calyculin A cell lysate <br />Lane 3: Rabbit IgG instead of <a href="/products/HA724447" style="font-weight: bold;text-decoration: underline;">HA724447</a> in NIH/3T3 treated with Calyculin A cell lysate <br /> <br />Exposure time: 11 seconds <br />Blocking: 5% NFDM/TBST, 1 hour at room temperature <br /> <br />Primary dilution: <a href="/products/HA724447" style="font-weight: bold;text-decoration: underline;">HA724447</a>, 1/2,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), 2 hours at room temperature <br /> <br />Predicted band size: 56 kDa <br />Observed band size: 56 kDa

    ☑ Cell treatment (CT)

    Immunoprecipitation (IP)

    Phospho-AKT (S473) was immunoprecipitated in 0.2 mg NIH/3T3 (Mouse fibroblast) treated with 100 nM Calyculin A for 30 minutes cell lysate with HA724447 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA724447 at 1/5,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature.

    Lane 1: NIH/3T3 treated with Calyculin A cell lysate (input)
    Lane 2: HA724447 IP in NIH/3T3 treated with Calyculin A cell lysate
    Lane 3: Rabbit IgG instead of HA724447 in NIH/3T3 treated with Calyculin A cell lysate

    Exposure time: 11 seconds
    Blocking: 5% NFDM/TBST, 1 hour at room temperature

    Primary dilution: HA724447, 1/2,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

    Predicted band size: 56 kDa
    Observed band size: 56 kDa

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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