bs-0237R [Primary Antibody]
Rabbit  Anti-14-3-3 Alpha + Beta + Gamma + Delta + Epsilon Rabbit pAb  Polyclonal Antibody
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Host: Rabbit

Target Protein: 14-3-3 Alpha + Beta + Gamma + Delta + Epsilon Rabbit pAb

IR: Immunogen Range:174-245/245

Clonality: Polyclonal

Isotype: IgG

Entrez Gene: 7529

Swiss Prot: P31946

Source: KLH conjugated synthetic peptide derived from human 14-3-3:174-245/245 

Purification: affinity purified by Protein A

Storage: 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20℃ for one year. Avoid repeated freeze/thaw cycles.

Background: 14-3-3 are activates tyrosine and tryptophan hydroxylases in the presence of Ca (2+)/calmodulin-dependent protein kinase II, and strongly activates protein kinase C. Is probably a multifunctional regulator of the cell signaling processes mediated by both kinases. Activates the ADP-ribosyltransferase (exoS) activity of bacterial origin. 14-3-3 proteins are localized in neurons, and are axonally transported to the nerve terminals. They may be also present, at lower levels, in various other eukaryotic tissues. It belongs to the 14-3-3 family.
This antibody is reactive with 14-3-3 Alpha, Beta, Gamma, Delta, Epsilon.

Size: 200ul

Concentration: 1mg/ml

Applications: WB=1:500-2000,IHC-P=1:100-500,IHC-F=1:100-500,IF=1:100-500,Flow-Cyt=1μg /test.

Cross Reactive Species: Human,Mouse,Rat (predicted: Sheep,Fruit Fly,Yeast)

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES
Sample: Cerebrum (Mouse) Lysate at 40 ug Primary: Anti-14-3-3 Alpha + Beta + Gamma + Delta + Epsilon (bs-0237R) at 1/300 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 27 kD Observed band size: 27 kD
Sample:Brain Cell Lysate at 40 ug Primary: Anti-14-3-3 Alpha + Beta + Gamma + Delta + Epsilon (bs-0237R) at 1/300 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 27 kD Observed band size: 27 kD
Sample: Lane 1: Cerebrum (Mouse) Lysate at 40 ug Lane 2: Uterus (Mouse) Lysate at 40 ug Lane 3: Testis (Mouse) Lysate at 40 ug Lane 4: Liver (Mouse) Lysate at 40 ug Lane 5: Lung (Mouse) Lysate at 40 ug Lane 6: Cerebrum (Rat) Lysate at 40 ug Lane 7: Uterus (Rat) Lysate at 40 ug Lane 8: Testis (Rat) Lysate at 40 ug Lane 9: Liver (Rat) Lysate at 40 ug Lane 10: Lung (Rat) Lysate at 40 ug Lane 11: NIH/3T3 (Mouse) Cell Lysate at 30 ug Lane 12: Siha (Human) Cell Lysate at 30 ug Lane 13: U251 (Human) Cell Lysate at 30 ug Primary: Anti-14-3-3 Alpha+Beta+Gamma+Delta+Epsilon (bs-0237R) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 28 kD Observed band size: 28 kD
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 (14-3-3 Alpha + Beta + Gamma + Delta + Epsilon) Polyclonal Antibody, Unconjugated (bs-1024R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse 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 (14-3-3 Alpha + Beta + Gamma + Delta + Epsilon) Polyclonal Antibody, Unconjugated (bs-1024R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Mouse 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 (14-3-3 Alpha + Beta + Gamma + Delta + Epsilon) Polyclonal Antibody, Unconjugated (bs-0237R) at 1:500 overnight at 4°C, followed by a conjugated secondary (sp-0023) for 20 minutes and DAB staining.
Tissue/cell: human rectal carcinoma;4% Paraformaldehyde-fixed and paraffin-embedded; Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min; Incubation: Anti-14-3-3 Polyclonal Antibody, Unconjugated(bs-0237R) 1:200, overnight at 4°C; The secondary antibody was Goat Anti-Rabbit IgG, PE conjugated(bs-0295G-PE)used at 1:200 dilution for 40 minutes at 37°C. DAPI(5ug/ml,blue,C-0033) was used to stain the cell nuclei
Blank control (blue line): A549 (blue). Primary Antibody (green line): Rabbit Anti-14-3-3 Alpha + Beta + Gamma + Delta + Epsilon antibody (bs-0237R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC Dilution: 1μg /test. Protocol The cells were fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% ice-cold methanol for 30 min on ice. Cells stained with Primary Antibody for 30 min at room temperature. The cells were then incubated in 1 X PBS/2%BSA/10% goat serum to block non-specific protein-protein interactions followed by the antibody for 15 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
Blank control:A431. Primary Antibody (green line): Rabbit Anti-14-3-3 Alpha + Beta + Gamma + Delta + Epsilon antibody (bs-0237R) Dilution: 1μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG . Secondary Antibody : Goat anti-rabbit IgG-AF647 Dilution: 1μg /test. Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1% PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min 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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