CoraLite® Plus 488-conjugated Acetyl-Tubulin (Lys40) Monoclonal antibody

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  IF Staining of mouse kidney using CL488-66200

  Frozen tissue section of adult mouse kidney was stained for acetylated ɑ-tubulin (magenta, Cat. No CL488-66200), CD-31/PECAM-1 (white), and Shh (green, Cat. No 20697-1-AP) with DAPI as a counterstain for visualizing the nucleus (blue). acetylated ɑ-tubulin stains primary cilia and was conjugated to Coralite-488 fluorescent dye and psedocolored to magenta. CD-31 stains endocardial/endothelial cells and was visualized with an Alexa Fluor 647 secondary antibody and psedocolored to white. Shh stains Sonic Hedgehog and was visualized with an Alexa Fluor 555 secondary antibody and psedocolored to green.The Image and figure legends are intellectual property of @LAF_in_the_LAB.

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  IF Staining of MDCK using CL488-66200

  Immunofluorescent analysis of (4% PFA) fixed MDCK cells using CL488-66200 (GREEN, Acetyl-Tubulin (Lys40) antibody) at dilution of 1:50 and CL594-17711 (RED, ARL13B antibody) at dilution of 1:50.

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  IF Staining of hTERT-RPE1 using CL488-66200

  Immunofluorescent analysis of (4% PFA) fixed hTERT-RPE1 cells using CoraLite® Plus 488 Acetyl-Tubulin (Lys40) antibody (CL488-66200, Clone: 7E5H8 ) at dilution of 1:200, CoraLite®594 ARL13B antibody (CL594-17711, red).

Acetyl-Tubulin (Lys40) Monoclonal Antibody for IF/ICC, IF-Fro

Cat No. CL488-66200

Clone No.7E5H8

Publications(5)

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat, pig, canine

Applications

IF/ICC, IF-Fro

Synonyms

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

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Tested Applications

• Positive IF-Fro detected in: mouse kidney tissue
• Positive IF/ICC detected in: MDCK cells, hTERT-RPE1 cells

Recommended dilution

• Immunofluorescence (IF)-FRO: IF-FRO : 1:50-1:500
• Immunofluorescence (IF)/ICC: IF/ICC : 1:50-1:500

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.

Published Applications

• IF: See 4 publications below

Product Information

CL488-66200 targets Acetyl-Tubulin (Lys40) in IF/ICC, IF-Fro applications and shows reactivity with human, mouse, rat, pig, canine samples.

• Tested Reactivity: human, mouse, rat, pig, canine
• Cited Reactivity: human, rat
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen:

  Peptide

• Full Name: tubulin, alpha 1a
• Calculated Molecular Weight: 52 kDa
• GenBank Accession Number: NM_006009
• Gene Symbol: Alpha Tubulin
• Gene ID (NCBI): 7846
• RRID: AB_2883282
• Conjugate: CoraLite® Plus 488 Fluorescent Dye
• Excitation/Emission Maxima Wavelengths: 493 nm / 522 nm
• Excitation Laser: Blue laser (488 nm)
• Form: Liquid
• Purification Method: Protein G purification
• UNIPROT ID: Q71U36
• Storage Buffer: PBS with 50% glycerol, 0.05% Proclin300, 0.5% BSA, pH 7.3.
• Storage Conditions: Store at -20°C. Avoid exposure to light. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage.

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).

Publications

• Human
• Mouse
• Rat
• Monkey
• Pig

Species	Application	Title
rat	IF	Front Cell Dev Biol; PIH1D3-knockout rats exhibit full ciliopathy features and dysfunctional pre-assembly and loading of dynein arms in motile cilia; Authors - Tingting Zhang
human	IF	STAR Protoc; Isolation, expansion, differentiation, and histological processing of human nasal epithelial cells; Authors - Vincent Manna
human	IF	BBA Adv; The dynamic nature of the coronavirus receptor, angiotensin-converting enzyme 2 (ACE2) in differentiating airway epithelia.; Authors - Vincent J Manna
		Cell Rep; Dairy cow- and avian-origin clade 2.3.4.4b H5N1 induce severe mastitis in lactating goats and transmission to suckling goats; Authors - Tamiru N Alkie
	IF	Am J Physiol Lung Cell Mol Physiol; Protocol For Differentiating Primary Human Small Airway Epithelial Cells at the Air-liquid Interface; Authors - Yu Par Aung Myo