Anti-Phospho-beta Actin (Tyr53) Polyclonal Antibody is a Rabbit antibody targeting Phospho-beta Actin (Tyr53). Anti-Phospho-beta Actin (Tyr53) Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat (predicted:Chicken,Pig,Cow,Rabbit)

1. Blank control: NIH/3T3.
Primary Antibody (green line): Rabbit Anti-phospho-beta Actin (Tyr53) antibody (TMAB-10385)
Dilution: 1 μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF488
Dilution: 1 μg /test.
Protocol
The cells were fixed with 4% PFA (10 min 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 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. Sample:
Lane 1: Hela (Human) Cell Lysate at 30 μg
Lane 2: MCF-7 (Human) Cell Lysate at 30 μg
Lane 3: Jurkat (Human) Cell Lysate at 30 μg
Primary: Anti-phospho-beta Actin (Tyr53) (TMAB-10385) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 42 kD
Observed band size: 42 kD
3. Sample: Pancreas (Mouse) Lysate at 40 μg
Primary: Anti-phospho-beta Actin (Tyr53) (TMAB-10385) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 42 kD
Observed band size: 42 kD
4. Paraformaldehyde-fixed, paraffin embedded (rat stomach 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 (beta Actin (Tyr53)) Polyclonal Antibody, Unconjugated (TMAB-10385) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated Synthesised phosphopeptide: human beta Actin around the phosphorylation site of Tyr53

Human

ACTB

Actin, cytoplasmic 1

Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.