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Home > Product > Antibody > Rabbit Anti-Phospho-SGK1 (Ser78) antibody
SGK1 (phospho S78); p-SGK1 (phospho S78); Serine/threonine protein kinase SGK; Serine/threonine protein kinase Sgk1; Serine/threonine-protein kinase Sgk1; Serum and glucocorticoid regulated kinase; Serum/glucocorticoid regulated kinase 1; Serum/glucocorti
Cat:
SL3395R
Species Reactivity：
Human,Mouse,Rat,(predicted: Dog,Pig,Cow,Horse,)
Immunogen：
KLH conjugated Synthesised phosphopeptide derived from human SGK1 around the phosphorylation site of Ser78:PP(p-S)PS
Format：
Liquid
Storage instructions：
Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
Concentration：
1mg/ml
Clonality：
Polyclonal
Isotype：
IgG
Applications：
WB=1:500-2000ELISA=1:5000-10000IHC-P=1:100-500IHC-F=1:100-500Flow-Cyt=2ug/TestIF=1:100-500（Paraffin sections need to do antigen repair）not yet tested in other applications.optimal dilutions/concentrations should be determined by the end user.
Host：
Rabbit
Product Overview：
Sample: A431 lysate at 30 ugPrimary: Rabbit Anti-Phospho-SGK1 (SL3395R) at 1:300 Secondary:IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size:49 kD Observed band size:49 kD Sample: A431 lysate at 30ugPrimary: Rabbit Anti-Phospho-SGK1 (SL3395R) at 1:300 Secondary: HRP conjugated Goat-Anti-rabbit IgG(SL0295G-HRP) at 1: 5000 Predicted band size:49 kD Observed band size:49 kD Sample: Pancreas (Mouse) Lysate at 40 ugPrimary: Anti-Phospho-SGK1 (Ser78) (SL3395R) at 1/1000 dilutionSecondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilutionPredicted band size: 49 kDObserved band size: 59 kDParaformaldehyde-fixed, paraffin embedded (mouse pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-SGK1 (Ser78)) Polyclonal Antibody, Unconjugated (SL3395R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (rat pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-SGK1 (Ser78)) Polyclonal Antibody, Unconjugated (SL3395R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-SGK1 (Ser78)) Polyclonal Antibody, Unconjugated (SL3395R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.Tissue/cell: human kidney tissue; 4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min; Incubation: Anti-Phospho-SGK1(Ser78) Polyclonal Antibody, Unconjugated(SL3395R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) stainingBlank control: Mouse spleen. Primary Antibody (green line): Rabbit Anti-SGK1 antibody (SL3395R) Dilution: 2μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-PEDilution: 1μg /test. ProtocolThe cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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Product PDFs
Datasheet:


SGK1 is a protein kinase that plays an important role in cellular stress response. SGK1 activates certain potassium, sodium, and chloride channels, suggesting an involvement in the regulation of processes such as cell survival, neuronal excitability, and renal sodium excretion. Sustained high levels of SGK1 and activity may contribute to conditions such as hypertension and diabetic nephropathy. This protein also mediates cell survival signals, as it has been shown to phosphorylate and negatively regulate the pro apoptotic FOXO3A protein. Ser 422 is a critical site on the protein and may be involved in its activation.

Function:
Protein kinase that plays an important role in cellular stress response. Activates certain potassium, sodium, and chloride channels, suggesting an involvement in the regulation of processes such as cell survival, neuronal excitability and renal sodium excretion. Sustained high levels and activity may contribute to conditions such as hypertension and diabetic nephropathy. Mediates cell survival signals, phosphorylates and negatively regulates pro-apoptotic FOXO3A. Phosphorylates NEDD4L, which leads to its inactivation and to the subsequent activation of various channels and transporters such as ENaC, KCNA3/Kv1.3 or EAAT1. Isoform 2 exhibited a greater effect on cell plasma membrane expression of ENaC and Na(+) transport than isoform 1.

Subunit:
Interacts with NEDD4 and NEDD4L.

Subcellular Location:
Isoform 2: Cell membrane.
Cytoplasm. Nucleus. Endoplasmic reticulum. Note=Nuclear, upon phosphorylation.

Tissue Specificity:
Expressed in most tissues with highest levels in the pancreas, followed by placenta, kidney and lung. Isoform 2 is strongly expressed in brain and pancreas, weaker in heart, placenta, lung, liver and skeletal muscle.

Post-translational modifications:
Regulated by phosphorylation. Phosphoinositide 3-kinase (PI3-kinase) pathway promotes phosphorylation at Ser-422 which in turn increases the phosphorylation of Thr-256 by PDPK1.
Ubiquitinated by NEDD4L; which promotes proteasomal degradation. Ubiquitinated by SYVN1 at the endoplasmic reticulum; which promotes rapid proteasomal degradation and maintains a high turnover rate in resting cells. Isoform 2 shows enhanced stability. Isoform 2 resistance to proteasomal degradation is mediated by the sequences within the first 120-amino acid.

Similarity:
Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family.
Contains 1 AGSLCkinase SLCterminal domain.
Contains 1 protein kinase domain.

SWISS:
O00141

Gene ID:
6446

Database links:

Entrez Gene: 6446 Human

Entrez Gene: 20393 Mouse

Entrez Gene: 29517 Rat

Omim: 602958 Human

SwissProt: O00141 Human

SwissProt: Q9WVC6 Mouse

SwissProt: Q06226 Rat

Unigene: 510078 Human

Unigene: 21685 Mouse

Unigene: 4636 Rat



Picture

Sample: A431 lysate at 30 ug
Primary: Rabbit Anti-Phospho-SGK1 (SL3395R) at 1:300
Secondary:IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size:49 kD
Observed band size:49 kD
Sample: A431 lysate at 30ug
Primary: Rabbit Anti-Phospho-SGK1 (SL3395R) at 1:300
Secondary: HRP conjugated Goat-Anti-rabbit IgG(SL0295G-HRP) at 1: 5000
Predicted band size:49 kD
Observed band size:49 kD
Sample:
Pancreas (Mouse) Lysate at 40 ug
Primary: Anti-Phospho-SGK1 (Ser78) (SL3395R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 49 kD
Observed band size: 59 kD
Paraformaldehyde-fixed, paraffin embedded (mouse pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-SGK1 (Ser78)) Polyclonal Antibody, Unconjugated (SL3395R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat pancreas); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-SGK1 (Ser78)) Polyclonal Antibody, Unconjugated (SL3395R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-SGK1 (Ser78)) Polyclonal Antibody, Unconjugated (SL3395R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Tissue/cell: human kidney tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-Phospho-SGK1(Ser78) Polyclonal Antibody, Unconjugated(SL3395R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining
Blank control: Mouse spleen.
Primary Antibody (green line): Rabbit Anti-SGK1 antibody (SL3395R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-PE
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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