Beta-Actin Loading Control Polyclonal Antibody 2 is a Rabbit antibody targeting Beta-Actin Loading Control. Beta-Actin Loading Control Polyclonal Antibody 2 can be used in FCM,IF,IHC-Fr,IHC-P,WB.

IgG

Human,Mouse,Rat (predicted:Pig,Cow,Sheep)

1. Sample:
Lane1: Hela (Human) Whole Cell Lysate at 30 μg
Lane2: Jurkat (Human) Whole Cell Lysate at 30 μg
Lane3: 293T (Human) Whole Cell Lysate at 30 μg
Lane4: Hepg2 (Human) Whole Cell Lysate at 30 μg
Lane5: A431 (Human) Whole Cell Lysate at 30 μg
Lane6: A549 (Human) Whole Cell Lysate at 30 μg
Lane7: H9C2 (Rat) Whole Cell Lysate at 30 μg
Lane8: Testis (Mouse) Tissue Lysate at 30 μg
Primary: Anti-F-Actin (TMAB-00217) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDa
2. Blank control (blue line): Hela (blue).
Primary Antibody (green line): Rabbit Anti-F-Actin antibody (TMAB-00217)
Dilution: 3 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.
Protocol
The cells were fixed with 80% methanol (5 min at-20°C) and then permeabilized with 0.1% PBS-Tween for 20 min at room temperature. 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.
3. Paraformaldehyde-fixed, paraffin embedded (rat stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (F-Actin) Polyclonal Antibody, Unconjugated (TMAB-00217) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (rat skeletal muscle); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (F-Actin) Polyclonal Antibody, Unconjugated (TMAB-00217) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

WB: 1:1000-5000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 3μg /test

Polyclonal

Synthetic MAP peptide: human beta-Actin

Human

ACTB

Actin, cytoplasmic 1

Actin,Actin,Beta Actin

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