Anti-Wnt5a Polyclonal Antibody is a Rabbit antibody targeting Wnt5a. Anti-Wnt5a Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,IF,FCM.

IgG

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

1. Tissue/cell: human lung carcinoma; 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-WNT5A Polyclonal Antibody, Unconjugated (TMAB-01986) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Formalin-fixed and paraffin embedded human breast labeled with Rabbit Anti-WNT5A Polyclonal Antibody (TMAB-01986) at 1:250 at room temperature overnight followed by conjugation to secondary antibody.
3. Sample: Ovary (Mouse) Lysate at 30 μg
Primary: Anti-WNT5A (TMAB-01986) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution
Predicted band size: 35 kDa
Observed band size: 47 kDa
4. Sample: Uters (Mouse) Lysate at 30 μg
Primary: Anti-WNT5A (TMAB-01986) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution
Predicted band size: 35 kDa
Observed band size: 47 kDa
5. Paraformaldehyde-fixed, paraffin embedded (rat ovary); 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 (WNT5A) Polyclonal Antibody, Unconjugated (TMAB-01986) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
6. Blank control: HepG2. Primary Antibody (green line): Rabbit Anti-WNT5A antibody (TMAB-01986)
Dilution: 1 μg/10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG.
Secondary Antibody: Goat anti-rabbit IgG-AF647
Dilution: 1 μg/test.
Protocol
The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 0.1% 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 room temperature. Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature.
7. Sample:
Lane 1: Embryo (Mouse) Lysate at 40 μg
Lane 2: Cerebrum (Mouse) Lysate at 40 μg
Lane 3: Uterus (Mouse) Lysate at 40 μg
Lane 4: Uterus (Rat) Lysate at 40 μg
Lane 5: Ovary (Rat) Lysate at 40 μg
Lane 6: Vas deferens (Rat) Lysate at 40 μg
Primary: Anti-WNT5A (TMAB-01986) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDa

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

Polyclonal

KLH conjugated synthetic peptide: human WNT5A

Human

PTC & Wnt pathway,Neural Signal Transduction,Secreted