Tubulin beta-III Recombinant Rabbit Monoclonal Antibody

• Catalog Number: orb612226
• Category: Antibodies
• Description: Tubulin beta-III Recombinant Rabbit Monoclonal Antibody
• Species/Host: Rabbit
• Clonality: Recombinant
• Clone Number: 3B7
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Mouse, Rat
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human Tubulin beta-III
• Antibody Type: Recombinant Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:1000-10000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100-500, IF=1:100-500, Flow-Cyt=1:50-200
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 50 kDa
• Target: TUBB3
• UniProt ID: Q13509
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: Neuron specific beta III Tubulin; beta 4; MC1R; TB
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Blank control: N2A. Primary Antibody (green line): Rabbit Anti-TUBB3 antibody (orb612226), dilution: 1:50, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF488, dilution: 1:1000. 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. Acquisition of 20000 events was performed.

Paraformaldehyde-fixed, paraffin embedded (human cerebellum), 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, Incubation with (Tubulin beta-III) Monoclonal Antibody, Unconjugated (orb612226) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Human left parietal lobe), 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, Incubation with (Tubulin beta-III) Monoclonal Antibody, Unconjugated (orb612226) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

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 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Incubation with (Tubulin beta-III) Monoclonal Antibody, Unconjugated (orb612226) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

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, Incubation with (Tubulin beta-III) Monoclonal Antibody, Unconjugated (orb612226) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (rat cerebellum), 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, Incubation with (Tubulin beta-III) Monoclonal Antibody, Unconjugated (orb612226) at 1:100 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Lane 1: Mouse Cerebrum tissue lysates, Lane 2: Mouse Cerebellum tissue lysates, Lane 3: Rat Cerebrum tissue lysates, Lane 4: Rat Cerebellum tissue lysates, Lane 5: Human SH-SY5Y cell lysates, Lane 6: Human U251 cell lysates, Lane 7: Human U87MG cell lysates, Primary: Anti-Tubulin beta-III (orb612226) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 50 kDa, Observed band size: 50 kDa.

SHG-44 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 (Tubulin beta-III (3B7)) Monoclonal Antibody, Unconjugated (orb612226) 1:50, 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.

SH-SY5Y 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 (Tubulin beta-III (3B7)) Monoclonal Antibody, Unconjugated (orb612226) 1:50, 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.