Beta III Tubulin Recombinant Rabbit Monoclonal Antibody [PSH15-32]
cat.: HA723738
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, IF-Tissue, IF-Cell, IP, IHC-Fr, FC(Intra)
Clonality: Monoclonal
Clone number: PSH15-32
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer: 1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Concentration: 0.1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 50 kDa
Isotype: IgG
Immunogen: Synthetic peptide within mouse Beta III Tubulin aa 401-450.
Positive control: SH-SY5Y cell lysate, U-87 MG cell lysate, Neuro-2a cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, mouse neuron cells, human cerebellum tissue, mouse E14.5 embryo tissue, mouse cerebellum tissue, rat E14.5 embryo tissue, rat cerebellum tissue, SH-SY5Y, Neuro-2a.
Subcellular location: Cytoplasm, cytoskeleton, growth cone, lamellipodium, filopodium.
Recommended Dilutions:
  WB
  IHC-P
  IF-Tissue
  IF-Cell
  IP
  IHC-Fr
  FC(Intra)
1:5,000
1:2,000-1:4,000
1:500-1:2,000
1:500-1:1,000
1-2μg/sample
1:500
1:1,000-1:10,000
Uniprot #: SwissProt: Q13509 Human | Q9ERD7 Mouse | Q4QRB4 Rat
Alternative names: beta 3 tubulin beta 4 beta-4 CDCBM CDCBM1 CFEOM3 CFEOM3A FEOM3 M(beta)3 M(beta)6 MC1R Neuron specific beta III Tubulin Neuron-specific class III beta-tubulin QccE-11995 QccE-15186 TBB3_HUMAN Tubb 3 TUBB3 TUBB4 Tubulin beta 3 Tubulin beta 3 chain Tubulin beta 4 Tubulin beta III Tubulin beta-3 chain Tubulin beta-4 chain Tubulin beta-III tuj 1 tuj1
Images
HA723738_1.jpg Fig1: Application: Immunofluorescence (IHC-Fr)

Species: Mouse
Tissue: Cerebellum
Sample: Frozen section

Antigen retrieval: Recommend. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.

Primary antibody: HA723738, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HA723738_2.jpg Fig2: Application: Immunofluorescence (IHC-Fr)

Species: Rat
Tissue: Cerebellum
Sample: Frozen section

Antigen retrieval: Recommend. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for about 2 minutes in microwave oven.

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.

Primary antibody: HA723738, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HA723738_3.jpg Fig3: Western blot analysis of Beta III Tubulin on different lysates with Rabbit anti-Beta III Tubulin antibody (HA723738) at 1/5,000 dilution.

Lane 1: SH-SY5Y (Human neuroblastoma cell) (15 µg/Lane)
Lane 2: HeLa (Human cervical adenocarcinoma cell) (15 µg/Lane)
Lane 3: U-87 MG (Human glioblastoma cell) (15 µg/Lane)
Lane 4: Neuro-2a (Mouse brain neuroblastoma cell) (15 µg/Lane)
Lane 5: NIH/3T3 (Mouse fibroblast) (15 µg/Lane)
Lane 6: Mouse brain tissue lysate (20 µg/Lane)
Lane 7: Rat brain tissue lysate (20 µg/Lane)

Exposure time: 4 seconds; ECL: K1801

HeLa is a low expression control (PMID: 12207023).

Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: HA723738, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

Predicted band size: 50 kDa
Observed band size: 50 kDa
HA723738_4.jpg Fig4: Application: Immunofluorescence (IF-tissue)

Species: Mouse
Tissue: Cerebellum
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: HA723738, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HA723738_5.jpg Fig5: Application: Immunofluorescence (IF-tissue)

Species: Rat
Tissue: Cerebellum
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.
Primary antibody: HA723738, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 1.5 hours at room temperature.
HA723738_6.jpg Fig6: Application: Immunocytochemistry (IF-cell)

Species: Mouse
Sample: Mouse neuron cells

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA723738, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
HA723738_7.jpg Fig7: Application: Immunohistochemistry (IHC-P)

Species: Human
Tissue: Cerebellum
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA723738, 1/4,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA723738_8.jpg Fig8: Application: Immunohistochemistry (IHC-P)

Species: Mouse
Tissue: E14.5 embryo
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA723738, 1/4,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA723738_9.jpg Fig9: Application: Immunohistochemistry (IHC-P)

Species: Mouse
Tissue: Cerebellum
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA723738, 1/4,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA723738_10.jpg Fig10: Application: Immunohistochemistry (IHC-P)

Species: Rat
Tissue: E14.5 embryo
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA723738, 1/4,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA723738_11.jpg Fig11: Application: Immunohistochemistry (IHC-P)

Species: Rat
Tissue: Cerebellum
Sample: Paraffin-embedded section

Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

Wash buffer: 1× TBST
Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
Primary antibody: HA723738, 1/4,000, 1 hour at room temperature.
Secondary antibody: HA1119, 20 minutes at room temperature.
HA723738_12.jpg Fig12: Application: Immunocytochemistry (IF-cell)

Species: Human
Sample: SH-SY5Y (Human neuroblastoma cell) / HeLa (Human cervical adenocarcinoma cell) (Low)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA723738, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

Low expression control: HeLa (PMID : 12207023).
HA723738_13.jpg Fig13: Application: Immunocytochemistry (IF-cell)

Species: Mouse
Sample: Neuro-2a (Mouse brain neuroblastoma cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.

Primary antibody: HA723738, 1/500, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue).
HA723738_14.jpg Fig14: Application: Flow Cytometry (Intra)

Species: Human
Sample: SH-SY5Y (Human neuroblastoma cell) / HeLa (Human cervical adenocarcinoma cell) (Low)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.

Primary antibody: HA723738 (1/10,000, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.

Low expression control: HeLa (PMID : 12207023).
HA723738_15.jpg Fig15: Application: Flow Cytometry (Intra)

Species: Mouse
Sample: Neuro-2a (Mouse brain neuroblastoma cell)

Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.

Primary antibody: HA723738 (1/1,000, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.
HA723738_16.jpg Fig16: Immunoprecipitation (IP)

Beta III Tubulin was immunoprecipitated in 0.2 mg mouse brain tissue lysate with HA723738 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA723738 at 1/5,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.

Lane 1: mouse brain tissue lysate (input)
Lane 2: HA723738 IP in mouse brain tissue lysate
Lane 3: Rabbit IgG instead of HA723738 in mouse brain tissue lysate

Exposure time: 7 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature

Primary dilution: HA723738, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature

Predicted band size: 50 kDa
Observed band size: 50 kDa
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.