Anticorps Monoclonal anti-acetylated Tubulin(Lys40)

acetylated Tubulin(Lys40) Monoclonal Antibody for IF, IHC, WB, ELISA

Hôte / Isotype

Mouse / IgG1

Réactivité testée

canin, Humain, porc, rat, souris et plus (3)

Applications

WB, IHC, IF, ELISA

Conjugaison

Non conjugué

Publications

75

CloneNo.

7E5H8

Avis (2)

N° de cat : 66200-1-Ig

Synonymes

Ac tubulin, acetylated alpha tubulin, Acetylated tubulin, Acetylated tubulin (Lys40), acetylated Tubulin(Lys40), acetylated-a-tubulin, acetyl-a-tubulin, Ac-tubulin, alpha tubulin, Alpha tubulin 3, TUBA1A, TUBA1A(ace-40Lys), TUBA3, tubulin, Tubulin alpha 3 chain, Tubulin B alpha 1, tubulin, alpha 1a

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Galerie de données de validation

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  WB analysis using 66200-1-Ig

  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.

• 

  WB analysis using 66200-1-Ig

  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.

• 

  WB analysis of pig cerebellum using 66200-1-Ig

  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.

• 

  WB analysis of rat brain using 66200-1-Ig

  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.

• 

  WB analysis using 66200-1-Ig

  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.

• 

  WB analysis of Neuro-2a using 66200-1-Ig

  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.

• 

  WB analysis of L02 using 66200-1-Ig

  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.

• 

  IHC staining of mouse ovary using 66200-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse ovary 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).

• 

  IHC staining of mouse brain using 66200-1-Ig

  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).

• 

  IHC staining of human gliomas using 66200-1-Ig

  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).

• 

  IHC staining of rat brain using 66200-1-Ig

  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).

• 

  IHC staining of human lung cancer using 66200-1-Ig

  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).

• 

  IF Staining of islet using 66200-1-Ig

  (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)

• 

  IF Staining of MDCK using 66200-1-Ig

  Immunofluorescent images of MDCK cells stained for IFT88 rabbit pAb (13967-1-AP) and acetylated tubulin mouse mAb (66200-1-Ig) at dilution of 1:50, further stained with Alexa Fluor 594-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) for 13967-1-AP, and Alexa Fluor 488-conjugated AffiniPure Goat anti-Mouse IgG (H+L) for 66200-1-Ig.

• 

  IF Staining of MDCK using 66200-1-Ig

  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).

• 

  IF Staining of mouse brain using 66200-1-Ig

  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).

• 

  ELISA experiment of Recombinant protein using 66200-1-Ig

  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.

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Applications testées

• Résultats positifs en WB: tissu testiculaire de rat, cellules HEK-293, cellules L02, cellules NCCIT, cellules Neuro-2a, testicule de souris, tissu cérébral de rat, tissu de cervelet de porc, tissu ovarien de souris
• Résultats positifs en IHC: tissu ovarien de souris, tissu cérébral de rat, tissu cérébral de souris, tissu de cancer du poumon humain, tissu de gliome humain; il est suggéré de démasquer l'antigène avec un tampon de TE buffer pH 9.0; (*) À défaut, 'le démasquage de l'antigène peut être 'effectué avec un tampon citrate pH 6,0.
• Résultats positifs en IF: cellules MDCK, tissu cérébral de souris
• Résultats positifs en ELISA: Protéine recombinante,

Dilution recommandée

• Western Blot (WB): WB : 1:2000-1:10000
• Immunohistochimie (IHC): IHC : 1:1000-1:4000
• Immunofluorescence (IF): IF : 1:50-1:500
• Test immuno-enzymatique (ELISA): ELISA : 1:10-1:100

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, check data in validation data gallery

Applications publiées

• WB: See 42 publications below
• IHC: See 2 publications below
• IF: See 39 publications below

Informations sur le produit

66200-1-Ig cible acetylated Tubulin(Lys40) dans les applications de WB, IHC, IF, ELISA et montre une réactivité avec des échantillons canin, Humain, porc, rat, souris

• Réactivité: canin, Humain, porc, rat, souris
• Réactivité citée: rat, bovin, Drosophile, Humain, poisson-zèbre, porc, souris
• Hôte / Isotype: Mouse / IgG1
• Clonalité: Monoclonal
• Type: Anticorps
• Immunogène: Peptide
• Nom complet: tubulin, alpha 1a
• Masse moléculaire calculée: 52 kDa
• Poids moléculaire observé: 50-55 kDa
• Numéro d’acquisition GenBank: NM_006009
• Symbole du gène: TUBA1A
• Identification du gène (NCBI): 7846
• Conjugaison: Non conjugué
• Forme: Liquide
• Méthode de purification: Purification par protéine G
• Tampon de stockage: PBS avec azoture de sodium à 0,02 % et glycérol à 50 % pH 7,3
• Conditions de stockage: Stocker à -20°C. Stable pendant un an après l'expédition. L'aliquotage n'est pas nécessaire pour le stockage à -20^oC Les 20ul contiennent 0,1% de BSA.

Informations générales

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).

Publications

Species	Application	Title
mouse	IF	Cell Stem Cell; Myoepithelial Cells of Submucosal Glands Can Function as Reserve Stem Cells to Regenerate Airways after Injury.; Authors - Aleksandra Tata
human	IF	J Clin Invest; Nasal ciliated cells are primary targets for SARS-CoV-2 replication in early stage of COVID-19.; Authors - Ji Hoon Ahn
mouse	WB	Aging Cell; SIRT2-knockdown rescues GARS-induced Charcot-Marie-Tooth neuropathy.; Authors - Yingying Zhao
	WB	Biomaterials; A virus-induced kidney disease model based on organ-on-a-chip: Pathogenesis exploration of virus-related renal dysfunctions.; Authors - Ji Wang
Drosophila	WB	Dev Cell; HDAC6 Suppresses Age-Dependent Ectopic Fat Accumulation by Maintaining the Proteostasis of PLIN2 in Drosophila.; Authors - Yan Yan
mouse	IF	Int J Biol Sci; NLRP3 Inflammasome Mediates Silica-induced Lung Epithelial Injury and Aberrant Regeneration in Lung Stem/Progenitor Cell-derived Organotypic Models; Authors - Hong Zhou

Avis

The reviews below have been submitted by verified Proteintech customers who received an incentive forproviding their feedback.

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FH

Mi (Verified Customer) (05-29-2023)

Excellent antibody, it works pretty well in human white adipocytes!

• Applications: Immunofluorescence
• Primary Antibody Dilution: 1:500
• Cell Tissue Type: adipocytes

FH

Sonia (Verified Customer) (04-08-2022)

Very good specificity and sensitivity. I used for IHC in different paraffin-embedded cell blocks for different cell lines, previously analyzed by WB.

• Applications: Immunohistochemistry
• Primary Antibody Dilution: 1:1000

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