CPT6 | cis-prenyltransferase 6

PRODUCT INFORMATION IN PDF

product information Background CPT6 (cis-prenyltransferase 6) belong to a group of enzymes which synthesize isoprenoid hydrocarbon skeleton with isoprenoid units in the cis (Z) configuration. AtCPT6 is one of the nine Arabidopsis thaliana CPTs, which catalyze the synthesis of a family of very short-chain polyisoprenoid alcohols of six, seven, and eight isoprenoid units. Immunogen KLH-conjugated synthetic peptide derived from Arabidopsis thaliana CPT6, UniProt:, Q8RX73, TAIR:AT5G58780 Host Rabbit Clonality Polyclonal Clone Purity Affinity purified serum Format Lyophilized in PBS pH 7.4 Quantity 50 µg Reconstitution For reconstitution add 50 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 Western blot (WB) Related products collection of antibodies to proteins involved in response to cold Plant and algal protein extraction buffer Secondary antibodies Additional information Surmacz described this protein in 20111 as AtCPT6. In TAIR is named ATCPT5 and in UniProt: Dehydrodolichyl diphosphate synthase 3.

application information
Recommended dilution		1 : 1000 (WB)
Expected | apparent MW		35 kDa
Confirmed reactivity		Arabidopsis thaliana
Predicted reactivity		please inquire
Not reactive in		No confirmed exceptions from predicted reactivity are currently known.
Additional information
Selected references		Surmacz et al. (2014). cis-Prenyltransferase AtCPT6 produces a family of very short-chain polyisoprenoids in planta. Biochim Biophys Acta. 2013 Dec 1;1841(2):240-250. doi: 10.1016/j.bbalip.2013.11.011.

application example


Microsomal (pellet P) and cytosolic (supernatant S) fractions from Arabidopsis thaliana roots were obtained by homogenization in homogenization buffer (50 mM Tris, pH 7.5, 5 mM MgCl2, 10 μM ZnCl2, 2 mM DTT, 100 mM NaCl, 250 mM sacharose) containing protease (Complete Mini, Roche) and phosphatase (PhosSTOP, Roche) inhibitor cocktails and centrifugation at 200,000 ×g for 1.5 h 25 µg of protein were separated on 12 % SDS-PAGE using wet transfer and blotted 1h to ECL nitrocellulose membrane. Blots were blocked with 4% non-fat milk in PBS-T (0.1% Tween-20 in 1× PBS) for 45 min at room temperature (RT) with agitation. Blot was incubated in the primary antibody 50 µg per 1 ml incubation mixture overnight at 4 °C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, and then washed once for 15 min and 3 times for 5 min in PBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit Gig horse radish peroxidase conjugated, from) diluted to 1:5 000 in for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturer's instructions. Exposure time was 2 minutes. S1 and S2 cytosolic (supernatant) and P1 and P2 microsomal (pellet) fractions were obtained from two independent experiments. Courtesy Dr. Liliana Surmacz, PAN Poland

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