Anti-Synaptopodin Polyclonal Antibody is a Rabbit antibody targeting Synaptopodin. Anti-Synaptopodin Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

Human,Mouse,Rat

1. 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 (Synaptopodin) Polyclonal Antibody, Unconjugated (TMAB-13318) at 1:400 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructions and DAB staining.
2. Positive control: RSC96
Isotype Control Antibody: Rabbit IgG;
Secondary Antibody: Goat anti-rabbit IgG-FITC;
Dilution: 1: 200 in 1 X PBS containing 0.5% BSA
Primary Antibody catalog number: TMAB-13318;
Dilution: 1 μg in 100 μl 1X PBS containing 0.5% BSA
3. Sample: Brain (Mouse) Lysate at 40 μg
Primary: Anti-Synaptopodin (TMAB-13318) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 102 kD
Observed band size: 102 kD
4. Sample: Cerebrum (Rat) Lysate at 40 μg
Primary: Anti-Synaptopodin (TMAB-13318) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 102 kD
Observed band size: 76 kD
5. Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by microwave in sodium citrate buffer (pH6.0); Block endogenous peroxidase by 3% hydrogen peroxide for 30 minutes; Blocking buffer (3% BSA) at RT for 30 min; Antibody incubation with (Synaptopodin) Polyclonal Antibody, Unconjugated (TMAB-13318) at 1:400 overnight at 4°C, followed by conjugation to the secondary antibody (labeled with HRP) and DAB staining.
6. Blank control (black line): sH-SY5Y.
Primary Antibody (green line): Rabbit Anti-Synaptopodin antibody (TMAB-13318)
Dilution: 1 μg/Test;
Secondary Antibody (white blue line): Goat anti-rabbit IgG-AF488
Dilution: 0.5 μg/Test.
Isotype control (orange line): Normal Rabbit IgG
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.

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

Polyclonal

KLH conjugated synthetic peptide: human Synaptopodin

Human

SYNPO

Synaptopodin