14-3-3 Alpha + Beta + Gamma + Delta + Epsilon Rabbit Polyclonal Antibody

• Catalog Number: orb13223
• Category: Antibodies
• Description: 14-3-3 Alpha + Beta + Gamma + Delta + Epsilon Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Drosophila, Sheep, Yeast
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human 14-3-3 (174-245/245aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1μg /test.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 27 kDa
• Target: YWHAB
• UniProt ID: P31946
• RRID: AB_10752379
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: YWHAB; YWHAZ; 14-3-3 alpha/beta/gamma/delta/Epsilo
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control (blue line): A549 (blue). Primary Antibody (green line): Rabbit Anti-14-3-3 Alpha + Beta + Gamma + Delta + Epsilon antibody (orb13223), Dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody (white blue line): Goat anti-rabbit IgG-FITC, Dilution: 1 µg/Test. Protocol, The cells were fixed with 70% ethanol (Overnight at 4°C) 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 1X 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 20000 events was performed.

Blank control: A431. Primary Antibody (green line): Rabbit Anti-14-3-3 Alpha + Beta + Gamma + Delta + Epsilon antibody (orb13223), Dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF647, Dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min 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 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (Mouse brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (14-3-3 Alpha + Beta + Gamma + Delta + Epsilon) Polyclonal Antibody, Unconjugated (orb13223) at 1:500 overnight at 4°C, followed by a conjugated secondary for 20 minutes and DAB staining.

Paraformaldehyde-fixed, paraffin embedded (mouse brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (14-3-3 Alpha + Beta + Gamma + Delta + Epsilon) Polyclonal Antibody, Unconjugated at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat brain), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (14-3-3 Alpha + Beta + Gamma + Delta + Epsilon) Polyclonal Antibody, Unconjugated at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Brain Cell Lysate at 40 ug, Primary: Anti-14-3-3 Alpha + Beta + Gamma + Delta + Epsilon (orb13223) 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: Cerebrum (Mouse) Lysate at 40 ug, Primary: Anti-14-3-3 Alpha + Beta + Gamma + Delta + Epsilon (orb13223) 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 (orb13223) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 28 kD, Observed band size: 28 kD.