Anti-Phospho-14-3-3 beta+zeta (Ser186/Ser184) Polyclonal Antibody is a Rabbit antibody targeting Phospho-14-3-3 beta+zeta (Ser186/Ser184). Anti-Phospho-14-3-3 beta+zeta (Ser186/Ser184) Polyclonal Antibody can be used in IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Chicken,Dog,Pig,Cow,Horse)

1. Blank control: A431.
Primary Antibody (green line): Rabbit Anti-phospho-14-3-3 beta+zeta (Ser186/Ser184) antibody (TMAB-10277)
Dilution: 2 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 20,000 events was performed.
2. Paraformaldehyde-fixed, paraffin embedded (rat brain tissue); 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 (YWHAB) Polyclonal Antibody, Unconjugated (TMAB-10277) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
3. Paraformaldehyde-fixed, paraffin embedded (mouse brain tissue); 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 (YWHAB) Polyclonal Antibody, Unconjugated (TMAB-10277) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
4. Blank control (blue line): HL60 (fixed with 70% ethanol (overninght at 4℃) and then permeabilized with 0.1% P Dilution: 0.2 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody (white blue line): Goat anti-rabbit IgG-PE, Dilution: 1 μg /test.

IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 0.2μg/Test

Polyclonal

KLH conjugated synthesised phosphopeptide: human 14-3-3 beta + zeta around the phosphorylation site of Ser186 / Ser184

Human

YWHAB

14-3-3 protein beta/alpha

Adapter protein implicated in the regulation of a large spectrum of both general and specialized signaling pathways. Binds to a large number of partners, usually by recognition of a phosphoserine or phosphothreonine motif. Binding generally results in the modulation of the activity of the binding partner.