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Beta III Tubulin Recombinant Antibody [PSH15-32] - Mouse IgG1 (Chimeric)

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Specification

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Catalog# HA601654

Beta III Tubulin Recombinant Antibody [PSH15-32] - Mouse IgG1 (Chimeric)

Application

  • IHC-P

  • IF-Tissue

  • IHC-Fr

  • WB

  • IF-Cell

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

Beta III Tubulin Recombinant Antibody [PSH15-32] - Mouse IgG1 (Chimeric)

Antibody Type

Recombinant Chimeric Antibody

Immunogen

Synthetic peptide within mouse Beta III Tubulin aa 401-450.

Species Reactivity

Human, Mouse, Rat

Validated Applications

IHC-P, IF-Tissue, IHC-Fr, WB, IF-Cell

Target Molecular Weight

Predicted band size: 50 kDa

Positive Control

Neuro-2a, Human Cerebellum, Mouse Cerebellum, Rat Cerebellum.

Conjugation

unconjugated

Clone Number

PSH15-32

Reactivity Data

IHC-P IF-Tissue IHC-Fr WB IF-Cell
Human
Tested Tested
Tested Tested
Tested Tested
Mouse
Tested Tested
Tested Tested
Tested Tested
Tested Tested
Tested Tested
Rat
Tested Tested
Tested Tested
Tested Tested

Product Features

Form

Liquid

Concentration

0.5 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

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.

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:10,000

  • IHC-P

  • 1:2,000

  • IF-Tissue

  • 1:1,000

  • IHC-Fr

  • 1:200-1:500

  • IF-Cell

  • 1:50

Target

Function

Tubulin beta-3 chain, Class III β-tubulin, βIII-tubulin (β3-tubulin) or β-tubulin III, is a microtubule element of the tubulin family found almost exclusively in neurons, and in testis cells. In humans, it is encoded by the TUBB3 gene. Forebrain neuronal culture after 40 days of differentiation from induced human pluripotent stem cells. Class III β-tubulin is one of the seven β-tubulin isotypes identified in the human genome, predominantly in neurons and the testis. It is conditionally expressed in a number of other tissues after exposure to a toxic microenvironment featured by hypoxia and poor nutrient supply. Posttranslational changes including phosphorylation and glycosylation are required for functional activity. Class III β-tubulin's role in neural development has warranted its use as an early biomarker of neural cell differentiation from multi potent progenitors. TUBB3 inactivation impairs neural progenitor proliferation. Rescue experiments demonstrate the non-interchangeability of TUBB3 with other classes of β-tubulins which cannot restore the phenotype resulting from TUBB3 inactivation. Congenital neurologic syndromes associated with TUBB3 missense mutations demonstrate the critical importance of class III β-tubulin for normal neural development.

Background References

1. Puri D et al. TUBB3 and KIF21A in neurodevelopment and disease. Front Neurosci. 2023 Aug

2. Jin S et al. TUBB3 M323V Syndrome Presents with Infantile Nystagmus. Genes (Basel). 2021 Apr

Subcellular Location

Cytoplasm, cytoskeleton, growth cone, lamellipodium, filopodium.

UNIPROT

SwissProt: Q13509 Human

SwissProt: Q9ERD7 Mouse

SwissProt: Q4QRB4 Rat

Synonyms

beta 3 tubulin antibody

beta 4 antibody

beta-4 antibody

CDCBM antibody

CDCBM1 antibody

CFEOM3 antibody

CFEOM3A antibody

FEOM3 antibody

M(beta)3 antibody

M(beta)6 antibody

Expand

Images

  • Application: Immunofluorescence (IHC-Fr)<br /><br />Species: Mouse<br />Tissue: Cerebral cortex<br />Sample: Frozen section<br /><br />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.<br /><br />Wash buffer: 1× TBST<br />Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.<br /><br />Primary antibody: <a href="/products/HA601654" style="font-weight: bold;text-decoration: underline;">HA601654</a>, 1/250, overnight at 4℃.<br />Secondary antibody: Goat Anti-Mouse IgG (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>), 1.5 hours at room temperature.

    Application: Immunofluorescence (IHC-Fr)

    Species: Mouse
    Tissue: Cerebral cortex
    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: HA601654, 1/250, overnight at 4℃.
    Secondary antibody: Goat Anti-Mouse IgG (iFluor™ 488, HA1125), 1.5 hours at room temperature.

  • Application: Immunofluorescence (IHC-Fr)<br /><br />Species: Rat<br />Tissue: Cerebral cortex<br />Sample: Frozen section<br /><br />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.<br /><br />Wash buffer: 1× TBST<br />Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature.<br /><br />Primary antibody: <a href="/products/HA601654" style="font-weight: bold;text-decoration: underline;">HA601654</a>, 1/250, overnight at 4℃.<br />Secondary antibody: Goat Anti-Mouse IgG (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>), 1.5 hours at room temperature.

    Application: Immunofluorescence (IHC-Fr)

    Species: Rat
    Tissue: Cerebral cortex
    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: HA601654, 1/250, overnight at 4℃.
    Secondary antibody: Goat Anti-Mouse IgG (iFluor™ 488, HA1125), 1.5 hours at room temperature.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Cerebellum<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/HA601654" style="font-weight: bold;text-decoration: underline;">HA601654</a>, 1/2,000, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1120" style="font-weight: bold;text-decoration: underline;">HA1120</a>, 20 minutes at room temperature.

    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: HA601654, 1/2,000, 1 hour at room temperature.
    Secondary antibody: HA1120, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Mouse<br />Tissue: Cerebellum<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/HA601654" style="font-weight: bold;text-decoration: underline;">HA601654</a>, 1/2,000, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1120" style="font-weight: bold;text-decoration: underline;">HA1120</a>, 20 minutes at room temperature.

    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: HA601654, 1/2,000, 1 hour at room temperature.
    Secondary antibody: HA1120, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Rat<br />Tissue: Cerebellum<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/HA601654" style="font-weight: bold;text-decoration: underline;">HA601654</a>, 1/2,000, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1120" style="font-weight: bold;text-decoration: underline;">HA1120</a>, 20 minutes at room temperature.

    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: HA601654, 1/2,000, 1 hour at room temperature.
    Secondary antibody: HA1120, 20 minutes at room temperature.

  • Application: Immunofluorescence (IF-tissue) <br /> <br />Species: Human <br />Tissue: Cerebellum <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature. <br />Primary antibody: <a href="/products/HA601654" style="font-weight: bold;text-decoration: underline;">HA601654</a>, 1/1,000, overnight at 4°C. <br />Secondary antibody: Goat Anti-Mouse IgG (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>), 1.5 hours at room temperature.

    Application: Immunofluorescence (IF-tissue)

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

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

    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: HA601654, 1/1,000, overnight at 4°C.
    Secondary antibody: Goat Anti-Mouse IgG (iFluor™ 488, HA1125), 1.5 hours at room temperature.

  • Application: Immunofluorescence (IF-tissue) <br /> <br />Species: Mouse <br />Tissue: Cerebellum <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature. <br />Primary antibody: <a href="/products/HA601654" style="font-weight: bold;text-decoration: underline;">HA601654</a>, 1/1,000, overnight at 4°C. <br />Secondary antibody: Goat Anti-Mouse IgG (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>), 1.5 hours at room temperature.

    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°C.

    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: HA601654, 1/1,000, overnight at 4°C.
    Secondary antibody: Goat Anti-Mouse IgG (iFluor™ 488, HA1125), 1.5 hours at room temperature.

  • Application: Immunofluorescence (IF-tissue) <br /> <br />Species: Rat <br />Tissue: Cerebellum <br />Sample: Paraffin-embedded section <br /> <br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95°C. <br /> <br />Wash buffer: 1× TBST <br />Blocking: 10% normal goat serum + 1% Triton X-100 + 0.3 M Glycine in TBST, 30 minutes at room temperature. <br />Primary antibody: <a href="/products/HA601654" style="font-weight: bold;text-decoration: underline;">HA601654</a>, 1/1,000, overnight at 4°C. <br />Secondary antibody: Goat Anti-Mouse IgG (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</a>), 1.5 hours at room temperature.

    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°C.

    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: HA601654, 1/1,000, overnight at 4°C.
    Secondary antibody: Goat Anti-Mouse IgG (iFluor™ 488, HA1125), 1.5 hours at room temperature.

  • Application: Immunocytochemistry (IF-cell) <br /> <br />Species: Human <br />Sample: Neuro-2a (Mouse brain neuroblastoma cell) <br /> <br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature. <br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature. <br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature. <br />Antibody dilution buffer: 1% BSA in PBST. <br /> <br />Primary antibody: <a href="/products/HA601654" style="font-weight: bold;text-decoration: underline;">HA601654</a>, 1/50, overnight at 4°C. <br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 45 minutes at room temperature. <br />Counterstain: Beta tubulin (<a href="/products/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, Red), 1/500, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    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: HA601654, 1/50, overnight at 4°C.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
    Counterstain: Beta tubulin (ET1602-4, Red), 1/500, overnight at 4℃. The nuclear counterstain was DAPI (Blue).

  • Western blot analysis of Beta III Tubulin on different lysates with Mouse anti-Beta III Tubulin antibody (<a href="/products/HA601654" style="font-weight: bold;text-decoration: underline;">HA601654</a>) at 1/10,000 dilution.<br /><br />Lane 1: SH-SY5Y (Human neuroblastoma cell)<br />Lane 2: HeLa (Human cervical adenocarcinoma cell)(Low expression)<br />Lane 3: Neuro-2a (Mouse brain neuroblastoma cell)<br />Lane 4: NIH/3T3 (Mouse fibroblast)(negative)<br /><br />Lysates/proteins at 10 µg/Lane.<br />Exposure time: 20 seconds; ECL: K1801<br /><br />HeLa is a relative low expression control (PMID: 35573698)<br />and NIH/3T3 is a negative control (PMID: 25839941).<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/HA601654" style="font-weight: bold;text-decoration: underline;">HA601654</a>, 1/10,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Mouse IgG-HRP (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 50 kDa<br />Observed band size: 50 kDa

    Western blot analysis of Beta III Tubulin on different lysates with Mouse anti-Beta III Tubulin antibody (HA601654) at 1/10,000 dilution.

    Lane 1: SH-SY5Y (Human neuroblastoma cell)
    Lane 2: HeLa (Human cervical adenocarcinoma cell)(Low expression)
    Lane 3: Neuro-2a (Mouse brain neuroblastoma cell)
    Lane 4: NIH/3T3 (Mouse fibroblast)(negative)

    Lysates/proteins at 10 µg/Lane.
    Exposure time: 20 seconds; ECL: K1801

    HeLa is a relative low expression control (PMID: 35573698)
    and NIH/3T3 is a negative control (PMID: 25839941).

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA601654, 1/10,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    Secondary antibody: Goat anti-Mouse IgG-HRP (HA1006), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 50 kDa
    Observed band size: 50 kDa

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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