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product information
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| background |
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Clathrin is a protein involved in intracellular trafficking and plays a major role in the formation of coated vesicles. It consists of three clathrin heavy chains and three light chains. Clathrin-coated vesicles (CCV) selectively sort cargo at the cell membrane, trans-Golgi network, and endosomal compartments for multiple membrane traffic pathways. |
| immunogen |
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KLH-conjugated peptide derived from available plant clathrin heavy chain sequences including Arabidopsis thaliana Q0WNJ6 |
| antibody format |
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rabbit |
polyclonal |
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affinity purified serum |
lyophilized |
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| quantity |
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100 µg |
for reconstitution add 100 µl of sterile water |
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| storage |
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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 |
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western blot (WB) |
| related products |
|
AS10 680 | Tubulin alpha chain AS10 681 | Tubulin beta chain |
| additional information |
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to be added when available |
application information
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| recommended dilution |
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1: 2000 with standard ECL (WB) |
| expected | apparent MW |
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193 | 170 kDa (Arabidopsis thaliana) |
| confirmed reactivity |
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Arabidopsis thaliana, Nicotiana tabacum
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| predicted reactivity |
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dicots including: Glycine max, Pisum sativum, Ricinus communis, monocots including: Oryza sativa, moss: Physcomitrella patens, algae: Chlamydomonas reinhardtii |
| not reactive in |
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no confirmed exceptions from predicted reactivity known in the moment |
| additional information |
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Please, note that fresh samples will provide better restuls (see image below). |
| selected references |
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to be added when available, antibody released in October 2011 |
application example
10 or 20 µl of freshly prepared total protein from Arabidopsis thaliana suspension culture (A), 10 or 20 µl of total protein (older extract) from Arabidopsis thaliana suspension culture (B), 10 or 20 µl of total leaf extract from Nicotiana tabacum (C), 10 or 20 µl of freshly prepared total protein from Nicotiana tabacum leaf protoplasts (D), 10 or 20 µl total protein from older extract of Nicotiana tabacum leaf protoplasts (E) , extracted with buffer containing 100 mM Tris (pH 7.8), 200 mM NaCl, 1 mM EDTA, 2% (v/v) beta-mercaptoethanol and 0.2% (v/v) Triton X-100, were separated on 8% SDS-PAGE and blotted 2h to nitrocellulose (semidry blot at 200 mA and RT). Blots were blocked with 5% (w/v) milk powder and 1% (w/v) BSA over night at 4°C with agitation. Blot was incubated in the primary antibody at a dilution of 1:2 000 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 Millipore) diluted to 1:10 000 in blocking solution for 45 min. 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 120 seconds.
Courtesy Fabian Künzl, University of Tuebingen, Germany.
||| For applications or usage on species others than stated as confirmed above Agrisera offers a payment-after-testing option. To learn more about this or for any questions on this product, please use the LiveChat option in the left menue bar or contact us at support@agrisera.com
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