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Tubulin alpha chain (monoclonal antibody)

AS20 4483  |  Clonality: Monoclonal  |  Host: Mouse |  Reactivity: Arabidopsis thaliana, Chlorella vulgaris, Eisenia, human, mouse, Nicotiana tabacum, Paramecium sp. , pig, turkey, yeast

Tubulin alpha chain (monoclonal antibody) in the group Antibodies for Plant/Algal  / Developmental Biology / Cytoskeleton at Agrisera AB (Antibodies for research) (AS20 4483)

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Product Information

Immunogen Fraction of tubulin purified from porcine brain by two cycles of polymerization - depolymerization, UniProt: Q71U36
Host Mouse
Clonality Monoclonal
Subclass/isotype IgG1
Purity Purified by Protein A affinity chromatography in PBS with 15 mM sodium azide.
Format Liquid
Quantity 100 µg
Storage Store at 4°C; Do not freeze.
Do not exceed exipry date is provided on the tube. 
Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Tested applications Immunocytochemistry (ICC), Immunohistochemistry on paraffin sections (IHC), Immunoprecipitation (IP), Western blot (WB)
Recommended dilution 1 : 500 (ICC), 1 : 1000 (WB)
Expected | apparent MW

51 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana, Chlorella vulgaris, Eisenia, human, mouse, Nicotiana tabacum, Paramecium sp. , pig, turkey, yeast
Predicted reactivity Plants
Species of your interest not listed? Contact us
Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application examples Western blot using monoclonal anti-tubulin alpha chain antibodies

Samples:

1 - 14 µg of Nicotiana tabacum, L. whole leaf extract of adult control plant (untreated)
2 - 14 µg of Nicotiana tabacum, L. whole leaf extract of adult plant after treatment with 100 µM AgNP-PVP
3 - 14 µg of Nicotiana tabacum, L. whole leaf extract of adult plant after treatment with 100 µM AgNP-CTAB
4 - 14 µg of Nicotiana tabacum, L. whole leaf extract of adult plant after treatment with 100 µM AgNO3
5 - 14 µg of Nicotiana tabacum, L. whole leaf extract of adult plant after treatment with 100 µM AgNP-PVP+cys
6 - 14 µg of Nicotiana tabacum, L. whole leaf extract of adult plant after treatment with 100 µM AgNP-CTAB+cys
7 - 14 µg of Nicotiana tabacum, L. whole leaf extract of adult plant after treatment with 100 µM AgNO3+cys
8 - 14 µg of Chlorella vulgaris extract- control (untreated)
9 – 7 µg of Arabidopsis thaliana seedlings extract- control (untreated)

14 µg/well of total protein extracted freshly from 0.05 g of lyophilized samples as specified above, following the Phenol extraction protocol 1 with extraction buffer containing Trizma base (500 mM), Ethylenediaminetetraacetic acid (EDTA) (50 mM), sucrose (700 mM) and Potassium chloride (KCl) (100 Mm) with addition of phenylmethylsulfonyl fluoride (PMSF) (1mM) and 2% β-mercaptoethanol. After short incubation on 4°C with agitation, phenol was added. The phenol (supernatant) phase containing proteins, was collected after centrifugation and equal volume of extraction buffer was added. After centrifugation, supernatant phase was collected and 4 volumes of 0.1 M ammonium acetate (with 10% methanol) was added, and proteins were precipitated ON/-20°C. The next day, protein pellets were washed 3 times in ammonium acetate with rounds of centrifugations in between, and finally in aceton. Protein pellet was lastly resuspended in Isoelectric focusing buffer (IEF) containing 9 M urea, 4% CHAPS, 20 mM DTT, 1.2% Ampholytes pH 3 to 10. Protein concentrations was measured with modified Bradford method1 and denatured with Laemmli sample buffer2 at 95°C for 5 min. Total proteins were separated on 12% SDS-PAGE and blotted 1h to nitrocellulose (pore size of 0.2 µm), using wet transfer. Blot was blocked with 2% milk in PBS-T, 1h/RT. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h/RT with agitation in a solution of 2% milk in PBS-T and then ON/4°C. The antibody solution was decanted and the blot was then washed 3 times for 10 min in 2% milk in PBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (goat anti-Mouse IgG1 HRP conjugated AS16 3715) diluted to 1:8000 in 2% milk in PBS-T for 1h/RT with agitation. The blot was washed twice for 10 min in PBS-T developed for 5 min with AgriseraECLSuperBright (AS16 ECL-S). Exposure time was 12 min.

Metal induced stress affected the expression of tubulin, and that therefore, this protein cannot be used as a loading control under that type of conditions

Courtesy of MSc, Karla Košpić, University of Zagreb, Faculty of Science Department of Biology, Croatia

Additional information

Additional information Metal induced stress affected the expression of tubulin, and that therefore, this protein cannot be used as a loading control under that type of conditions, data in application example. 
This antibody is recognizing defined epitope (amino acid 65-97) on N-terminal structural domain of alpha tubulin.

Recommended secondary antibody, goat anti-mouse IgG1, HRP conjugated AS16 3715

Related products

Related products
Collection of antibodies to cytoskeleton proteins
AS10 680 | Anti-tubulin alpha chain, rabbit anitbodies
AS10 681 | Anti-tubulin beta chain, rabbit antibodies

Background

Background Tubulin alpha (TUA) together with beta tubulin is making up microtubules.

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