TUBB3 (Neuronal Marker) Rabbit Polyclonal Antibody

• Catalog Number: orb500814
• Category: Antibodies
• Description: TUBB3 (Neuronal Marker) Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Canine, Rabbit
• Reactivity: Human, Mouse, Rat
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human beta III Tubulin (401-450/450aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:200-800, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 50-55 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 (blue): U-87MG Cells (fixed with 2% paraformaldehyde (10 min)). Primary Antibody: Rabbit Anti-MGLUR3/AF647 Conjugated antibody, dilution: 1 µg in 100 µl 1X PBS containing 0.5% BSA, Isotype Control Antibody: Rabbit IgG/AF647 (orange), used under the same conditions.

Blank control: SH-SY5Y. Primary Antibody (green line): Rabbit Anti-TUBB3 (Neuronal Marker) antibody (orb500814), dilution: 1 ug/Test, Secondary Antibody: Goat anti-rabbit IgG-FITC, dilution: 0.5 ug/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. Acquisition of 20000 events was performed.

Blank control: SH-SY5Y. Primary Antibody (green line): Rabbit Anti-TUBB3 (Neuronal Marker) antibody (orb500814), dilution: 1 ug/Test, Secondary Antibody: Goat anti-rabbit IgG-FITC, dilution: 0.5 ug/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. Acquisition of 20000 events was performed.

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 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (TUBB3) Polyclonal Antibody, Unconjugated (orb500814) at 1:400 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody for 90 minutes, and DAPI for nuclei 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, Antibody incubation with (TUBB3 (Neuronal Marker)) Polyclonal Antibody, Unconjugated (orb500814) at 1:200 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, Antibody incubation with (TUBB3) Polyclonal Antibody, Unconjugated (orb500814) at 1:400 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody for 90 minutes, and DAPI for nuclei staining.

Sample: BV-2 (Rat) Cell Lysate at 30 ug, Primary: Anti-TUBB3 (Neuronal Marker) (orb500814) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 50-55 kD, Observed band size: 50 kD.

Sample: U251 (Human) Cell Lysate at 30 ug, Primary: Anti-TUBB3 (Neuronal Marker) (orb500814) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 50-55 kD, Observed band size: 50 kD.

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 (orb500814) 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 nuclei.

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 (orb500814) 1:200, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (orb868805) at 37°C for 90 minutes, DAPI (5 ug/ml, blue) was used to stain the cell nuclei.