Acetyl-Tubulin (Lys40) Monoclonal antibody, PBS Only

Acetyl-Tubulin (Lys40) Monoclonal Antibody for WB, IHC, IF/ICC, ELISA, Indirect ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat, pig, canine

Applications

WB, IHC, IF/ICC, ELISA, Indirect ELISA

Conjugate

Unconjugated

CloneNo.

7E5H8

Cat no : 66200-1-PBS

Synonyms

acetylated alpha tubulin, TUBA1A, Tubulin alpha 1A chain, 7E5H8, Detyrosinated tubulin alpha-1A chain

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Validation Data Gallery

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  WB analysis using 66200-1-Ig (same clone as 66200-1-PBS)

  Various lysates were subjected to SDS PAGE followed by western blot with 66200-1-Ig (Acetyl-Tubulin (Lys40) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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  WB analysis using 66200-1-Ig (same clone as 66200-1-PBS)

  Mouse ovary lysates (Left lane: 40 μg, Right lane: 20 μg) were subjected to SDS PAGE followed by western blot with 66200-1-Ig (acetylated Tubulin(Lys40) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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  WB analysis of pig cerebellum using 66200-1-Ig (same clone as 66200-1-PBS)

  pig cerebellum tissue were subjected to SDS PAGE followed by western blot with 66200-1-Ig (acetylated Tubulin(Lys40) antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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  WB analysis of rat brain using 66200-1-Ig (same clone as 66200-1-PBS)

  rat brain tissue were subjected to SDS PAGE followed by western blot with 66200-1-Ig (acetylated Tubulin(Lys40) antibody) at dilution of 1:20000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

• 

  WB analysis using 66200-1-Ig (same clone as 66200-1-PBS)

  Various lysates were subjected to SDS PAGE followed by western blot with 66200-1-Ig (acetylated Tubulin(Lys40) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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  WB analysis of Neuro-2a using 66200-1-Ig (same clone as 66200-1-PBS)

  Neuro-2a cells were subjected to SDS PAGE followed by western blot with 66200-1-Ig (acetylated Tubulin(Lys40) antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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  WB analysis of L02 using 66200-1-Ig (same clone as 66200-1-PBS)

  L02 cells were subjected to SDS PAGE followed by western blot with 66200-1-Ig (acetylated Tubulin(Lys40) antibody at dilution of 1:1000 incubated at room temperature for 1.5 hours. This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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  IHC staining of mouse ovary using 66200-1-Ig (same clone as 66200-1-PBS)

  Immunohistochemical analysis of paraffin-embedded mouse ovary tissue slide using 66200-1-Ig (Acetyl-Tubulin (Lys40) antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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  IHC staining of mouse brain using 66200-1-Ig (same clone as 66200-1-PBS)

  Immunohistochemical analysis of paraffin-embedded mouse brain tissue slide using 66200-1-Ig (acetylated Tubulin(Lys40) antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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  IHC staining of human gliomas using 66200-1-Ig (same clone as 66200-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human gliomas tissue slide using 66200-1-Ig (acetylated Tubulin(Lys40) antibody) at dilution of 1:2000 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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  IHC staining of human lung cancer using 66200-1-Ig (same clone as 66200-1-PBS)

  Immunohistochemical analysis of paraffin-embedded human lung cancer tissue slide using 66200-1-Ig (acetylated Tubulin(Lys40) antibody at dilution of 1:50 (under 10x lens). This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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  IHC staining of rat brain using 66200-1-Ig (same clone as 66200-1-PBS)

  Immunohistochemical analysis of paraffin-embedded rat brain tissue slide using 66200-1-Ig (acetylated Tubulin(Lys40) antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0). This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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  IF Staining of islet using 66200-1-Ig (same clone as 66200-1-PBS)

  (left) Confocal image of a single-plane cross-section of a healthy human islet, showing primary cilia on both alpha cells (glucagon, red) and non-alpha cells (non-red). Acetylated alpha tubulin, cilia (green, Cat. No 66200-1-Ig, 1:400), nuclei (blue), scale 20 μm. (right) Primary cilia in wildtype B6 mouse islets: acetylated alpha tubulin (green, Cat. No 66200-1-Ig, 1:400), glucagon (red), nuclei (blue), single-plane images, scales 10 μm. (Fig from Dr. Jing Hughes) This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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  IF Staining of mouse brain using 66200-1-Ig (same clone as 66200-1-PBS)

  Immunofluorescent analysis of (4% PFA) fixed mouse brain tissue using 66200-1-Ig (acetylated Tubulin(Lys40) antibody) at dilution of 1:100 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG(H+L). This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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  IF Staining of hTERT-RPE1 using 66200-1-Ig (same clone as 66200-1-PBS)

  Immunofluorescent analysis of (4% PFA) fixed hTERT-RPE1 cells using acetylated Tubulin(Lys40) antibody (66200-1-Ig, Clone: 7E5H8 ) at dilution of 1:800 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L), CoraLite®594 ARL13B antibody (CL594-17711, red). This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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  IF Staining of MDCK using 66200-1-Ig (same clone as 66200-1-PBS)

  Immunofluorescent analysis of MDCK cells using 66200-1-Ig (acetylated Tubulin (Lys40) antibody) at dilution of 1:100 and Alexa Fluor 488-conjugated AffiniPure Goat Anti-Mouse IgG (H+L). This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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  ELISA experiment of Recombinant protein using 66200-1-Ig (same clone as 66200-1-PBS)

  Specificity validation by indirect ELISA. Indirect ELISA was carried out by coating E.coli expressed GST-tagged Tubulin alpha (70 ng/well), non-acetylated Tubulin alpha, and Lys40 acetylated Tubulin alpha (with excessive amount) followed by blocking and adding serial diluted GST tag primary antibody (Cat.NO. 66001-2-Ig) and Acetylated Tubuliln antibody 66200-1-Ig respectively. HRP-Goat anti-mouse secondary antibody was used for detection. Signal was developed with TMB and stopped by H2SO4. Signal strength was measured by absorbance at 450 nm. This data was developed using the same antibody clone with 66200-1-PBS in a different storage buffer formulation.

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Product Information

66200-1-PBS targets Acetyl-Tubulin (Lys40) in WB, IHC, IF/ICC, ELISA, Indirect ELISA applications and shows reactivity with human, mouse, rat, pig, canine samples.

• Tested Reactivity: human, mouse, rat, pig, canine
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: Peptide
• Full Name: tubulin, alpha 1a
• Calculated Molecular Weight: 52 kDa
• Observed Molecular Weight: 50-55 kDa
• GenBank Accession Number: NM_006009
• Gene Symbol: Alpha Tubulin
• Gene ID (NCBI): 7846
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS Only
• Storage Conditions: Store at -80°C.

Background Information

What is the molecular weight of acetyl-α-tubulin?

The molecular weight of acetylated tubulin is 52 kD.

Where does the acetylation of Lys 40 occur?

This acetylation occurs inside the microtubule lumen by the α-tubulin acetyltransferase 1 (αTAT1) (PMID: 29207274).

How can acetylation be reversed?

Acetylation of Lys 40 can be reversed by deacetylase 6 (HDAC6), which is mostly cytoplasmic that also deacetylates Hsp90, and sirtuin 2 (SIRT2), which also mainly cytoplasmic and uses NAD as a coenzyme. Unlike HDAC6, SIRT uses both polymerized and soluble tubulin as substrates. Deacetylases are believed to be more active on soluble tubulin, while acetylases function preferentially on stable polymers (PMIDs: 29207274, 30079247, 19185337).

What is the function of acetylation?

Acetylation is a conserved post-translational modification of alpha tubulin at Lys 40 during tubulin assembly, and it correlates to increased microtubule stability and intracellular transport (PMIDs: 29207274, 30079247, 20940043).

Is acetylation of α-tubulin strictly associated with stable microtubules?

Not necessarily, as acetylation can have other effects on microtubule subpopulations (PMID 20940043).

Is ac-tubulin found only in cilia?

Acetylated-α-tubulin is located in cytoplasmic tubulin as well as in cilia; therefore, it is not strictly region-specific (PMID: 30079247).

What are the cellular effects of tubulin acetylation?

Microtubule acetylation seems to provide a critical role in neuronal development and function, and while its effect on cancer cells remains unclear, it has been shown that decreased acetylated α-tubulin impairs neuronal cell line migration. The post-translational modification may also help regulate organelle-independent signalling throughout the cell, supporting the notion of a microtubule network serving as a coordinator of cellular signaling (PMIDs: 29207274, 25503560, 20940043, 19185337).