Anti-VAX1 Polyclonal Antibody is a Rabbit antibody targeting VAX1. Anti-VAX1 Polyclonal Antibody can be used in WB,ICC/IF,FCM.

IgG

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

1. A549 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 (VAX1) polyclonal Antibody, Unconjugated (TMAB-14058) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.
2. Sample: BRL-3A Cell (Rat) Lysate at 40 μg
Primary: Anti-VAX1 (TMAB-14058) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 35 kD
Observed band size: 37 kD
3. Sample: NIH/3T3 Cell (Mouse) Lysate at 40 μg
Primary: Anti-VAX1 (TMAB-14058) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 35 kD
Observed band size: 37 kD
4. Blank control (blue line): Hep G2 (fixed with 70% ethanol (Overnight at 4℃) and then permeabilized with 90% methanol for 20 min at -20℃).
Primary Antibody (green line): Rabbit Anti-VAX1 antibody (TMAB-14058), Dilution: 0.2 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.
5. Sample:
MCF-7 Cell (Human) Lysate at 40 μg
U937 Cell (Human) Lysate at 40 μg
Primary: Anti-VAX1 (TMAB-14058) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 35 kD
Observed band size: 36 kD
6. Blank control: A431.
Primary Antibody (green line): Rabbit Anti-VAX1 antibody (TMAB-14058)
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 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.
7. Blank control (black line): A549.
Primary Antibody (green line): Rabbit Anti-VAX1 antibody (TMAB-14058)
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; ICC/IF: 1:100-500; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human VAX1

Human

VAX1

Ventral anterior homeobox 1

Required for axon guidance and major tract formation in the developing forebrain. May contribute to the differentiation of the neuroretina, pigmented epithelium and optic stalk.