Anti-TUBB3 Polyclonal Antibody is a Rabbit antibody targeting TUBB3. Anti-TUBB3 Polyclonal Antibody can be used in WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM.

IgG

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

1. Tissue/cell: rat brain 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-TUBB3/beta III Tubulin (Neuronal Marker) Polyclonal Antibody, Unconjugated (TMAB-01919) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
2. Tissue/cell: BV-2 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 (TUBB3) Polyclonal Antibody, Unconjugated (TMAB-01919) 1:200, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (5 μg/ml, blue) was used to stain the cell nucleus.
3. Blank control (blue): U-87MG Cells (fixed with 2% paraformaldehyde (10 min)). P
rimary Antibody: Rabbit Anti-MGLUR3/AF647 Conjugated antibody (TMAB-01919/AF647), Dilution: 1 μg in 100 μL 1X PBS containing 0.5% BSA;
Isotype Control Antibody: Rabbit IgG/AF647 (orange),used under the same conditions.
4. Paraformaldehyde-fixed, paraffin embedded (mouse cerebellum); 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 (TUBB3 (Neuronal Marker)) Polyclonal Antibody, Unconjugated (TMAB-01919) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
5. SH-SY5Y cell; 4% Paraformaldehyde-fixed; Ice-cold methanol at-20°C for 20 min; Blocking buffer (normal goat serum) at 37°C for 20 min; Antibody incubation with (TUBB3) polyclonal Antibody, Unconjugated (TMAB-01919) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nucleus.
6. Blank control: SH-SY5Y.
Primary Antibody (green line): Rabbit Anti-TUBB3 (Neuronal Marker) antibody (TMAB-01919)
Dilution: 1 μg/Test;
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 0.5 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°C. 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. Blank control: SH-SY5Y.
Primary Antibody (green line): Rabbit Anti-TUBB3 (Neuronal Marker) antibody (TMAB-01919)
Dilution: 1 μg/Test;
Secondary Antibody: Goat anti-rabbit IgG-FITC
Dilution: 0.5 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°C. 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.
8. 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 (TUBB3) Polyclonal Antibody, Unconjugated (TMAB-01919) at 1:400 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody for 90 minutes, and DAPI for nucleus staining.
9. Paraformaldehyde-fixed, paraffin embedded (Mouse 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 (TUBB3) Polyclonal Antibody, Unconjugated (TMAB-01919) at 1:400 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody for 90 minutes, and DAPI for nucleus staining.
10. Sample:
U251 (Human) Cell Lysate at 30 μg
Primary: Anti-TUBB3 (Neuronal Marker) (TMAB-01919) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 50-55 kDa
Observed band size: 50 kDa
11. Sample:
BV-2 (Rat) Cell Lysate at 30 μg
Primary: Anti-TUBB3 (Neuronal Marker) (TMAB-01919) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 50-55 kDa
Observed band size: 50 kDa

WB,IHC-P,IHC-Fr,ICC/IF,IF,FCM

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; ICC/IF: 1:100; IF: 1:200-800; FCM: 1μg/Test

Polyclonal

KLH conjugated synthetic peptide: human beta III Tubulin

Human

Neuronal,Tubulin,Microtubules