Anti-SM22 Alpha Polyclonal Antibody is a Rabbit antibody targeting SM22 Alpha. Anti-SM22 Alpha Polyclonal Antibody can be used in FCM, ICC/IF, IF, IHC-Fr, IHC-P.

IgG

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

1. Tissue/cell: U251 cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (SM22 Alpha) Polyclonal Antibody, Unconjugated (TMAB-01733) 1:200, 90 minutes at 37°C; followed by a conjugated secondary antibody at 37°C for 90 minutes, DAPI (5 μg/ml, blue) was used to stain the cell nucleus.
2. Blank control: Raji.
Primary Antibody (green line): Rabbit Anti-SM22 Alpha antibody (TMAB-01733)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-PE
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 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 at room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
3. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (SM22 Alpha) Polyclonal Antibody, Unconjugated (TMAB-01733) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
4. Paraformaldehyde-fixed, paraffin embedded (Rat bladder); 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 (SM22 Alpha) Polyclonal Antibody, Unconjugated (TMAB-01733) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
5. Paraformaldehyde-fixed, paraffin embedded (Human uterus); 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 (SM22 Alpha) Polyclonal Antibody, Unconjugated (TMAB-01733) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
6. Paraformaldehyde-fixed, paraffin embedded (Rat heart); 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 (SM22 Alpha) Polyclonal Antibody, Unconjugated (TMAB-01733) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
7. Paraformaldehyde-fixed, paraffin embedded (mouse heart); 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 (SM22 Alpha) Polyclonal Antibody, Unconjugated (TMAB-01733) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

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

Polyclonal

KLH conjugated synthetic peptide: human SM22 Alpha

Human

TAGLN

Transgelin

Smooth Muscle Cells,Inotropics,Zinc Finger,Actin Binding Proteins,Calcium Signaling

Actin cross-linking/gelling protein. Involved in calcium interactions and contractile properties of the cell that may contribute to replicative senescence.