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Tubulin alpha chain

265 €
Buy 2 items of this product for 198 €/each
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AS10 680  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: Arabidopsis thaliana, Euglena gracilis, Horderum vulgare, Zea mays

PRODUCT INFORMATION IN PDF

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AS10 680

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product information
Background

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

Immunogen

KLH-conjugated peptide derived from available tubulin alpha chain sequences including Arabidopsis thaliana tubulin alpha-1-chain P11139(At1g64740), alpha-2/alpha-4 chain B9DGT7(At1g50010), alpha-5 chain B9DHQ0(At5g19780), alpha-6-chain P29511(At4g14960)

Host Rabbit
Clonality Polyclonal
Clone
Purity Serum
Format Lyophilized
Quantity 100 µl
Reconstitution
For reconstitution add 100 µl of sterile water
Storage

Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. 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 Immunofluorescence (IF), Western Blot (WB)
Related products
AS10 681 | tubulin beta chain
Additional information

Please, note that tubulin alpha is only detected in actively dividing meristematic cells (see immunolocalization image below). Therefore to unable detection on a western blot, analyzed material must contain enough cells with dividing activity.

application information
Recommended dilution

1: 1000 with standard ECL (WB), 1: 500 (IF)

Expected | apparent MW

49 | 52 kDa (Arabidopsis thaliana)

Confirmed reactivity

Arabidopsis thaliana, Euglena gracilis, Hordeum vulgare, Zea mays

Predicted reactivity

dicots including: Brassica napus, Glycine max, Pisum sativum, Solanum tuberosum, Sorghum bicolor, Ricinus communis, Vitis vinifera, monocots including: Triticum aestivum, Oryza sativa, trees: Picea sitchensis, Populus trichocarpa, moss: Physcomitrella patens, algae: Chlorella vulgaris, Euglena gracilis, Micromonoas pusilla,  Ostreococcus lucimarinus

Not reactive in

no confirmed exceptions from predicted reactivity known in the moment

Additional information

Peptide used to elict this antibody is not present in tubulin beta.

Selected references
Nasir (2016). Analysis of signal transduction chains of gravity and light sensing in Euglena gracilis. Doctoral Thesis, urn:nbn:de:bvb:29-opus4-79185
Juszczak et al. (2012). Natural genetic variation in the expression regulation of the chloroplast antioxidant system among Arabidopsis thaliana accessions. Physiol. Plant.

Application example

Western Blot

western blot using anti-plant tubulin alpha antibody

 5 µg of total protein from Arabidopsis thaliana (1), Hordeum vulgare (2), Zea mays (3)   extracted with Agrisera PEB extraction buffer were separated on  4-12 % SDS-PAGE and blotted 1h to PVDF. Blots were blocked with   for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 10 000 for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602 ) diluted to 1:25 000 in  for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL Advance according to the manufacturers instructions (GE Health care). Exposure time was 120 seconds.

western blot using anti-tubulin alpha antibody on rice embryo

Total protein from either rice embryos or rice protoplasts extracted with buffer containing 60mM Tris-HCl (pH8.0), 2% SDS(w/v), 15% Sucrose (w/v) and protease inhibitor 1X were separated on 12 % SDS-PAGE and blotted 1h to PVDF. Blots were blocked with skimmed milk containing TBS 1X for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 overnight at 4 degree with shaking about 40 rpm. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:20 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturers instructions. Exposure time was 10 seconds.

Courtesy of Ho Viet The, PhD Student, Scuola Superiore Sant'Anna (Pisa, Italy).

Immunolocalization

immunolocalization of plant tubulin alpha chain

Tubulin alpha localization in roots of Arabidopsis thaliana.  Tubulin alpha (red), nucleus (DAPI white). Plant material has been fixed in para-formaldehyde for 30 minutes. Tissue cleaning has been performed before immunolocalization. Primary antibodies: Agrisera anti-tubulin alpha 1: 500. Secondary antibody:  goat anti-rabbit IgG Alexa conjugated (red color), dilution 1: 500. Scale bar – 10 µm.

Courtesy Dr. Taras Pasternak, Freiburg University, Germany


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