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

IgG

Human,Mouse,Rat,Dog (predicted:Rabbit)

1. Sample:
MDCk (Dog) Cell Lysate at 40 μg
Primary: Anti-Nephrin (TMAB-01214) at 1/500 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 138 kDa
Observed band size: 138 kDa
2. Sample:
Liver (Mouse) Lysate at 40 μg
Primary: Anti-Nephrin (TMAB-01214) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 138 kDa
Observed band size: 138 kDa
3. Blank control: 293T.
Primary Antibody (green line): Rabbit Anti-Nephrin antibody (TMAB-01214)
Dilution: 1 μg/Test;
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 0.5 μg/Test.
Protocol
The cells wereincubated 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.
4. Tissue/cell: mouse kidney tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), Boiling bathing for 15 min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min; Blocking buffer (normal goat serum) at 37°C for 20 min;
Incubation: Anti-Nephrin Polyclonal Antibody, Unconjugated (TMAB-01214) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
5. the cells (293T) were fixed with 2% paraformaldehyde (10 min).
Isotype Control Antibody: Rabbit Igg (orange);
Secondary Antibody: Goat anti-rabbit IgG-Pe (white blue),
Dilution: 1:200 in 1 X PBS containing 0.5% BSA;
Primary Antibody Dilution: 1 μg in 100 μL 1X PBS containing 0.5% BSA (green).
6. Blank control: RSC96 (blue), the cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with ice-cold 90% methanol for 30 min on ice.
Isotype Control Antibody: Rabbit Igg (orange);
Secondary Antibody: Goat anti-rabbit IgG-Pe (white blue),
Dilution: 1:200 in 1 X PBS containing 0.5% BSA;
Primary Antibody Dilution: 1 μg in 100 μL 1X PBS containing 0.5% BSA (green).

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 Nephrin

Human

Endothelial