Acd1 | accelerated cell death 1

315 €

AS11 1783 | clonality: polyclonal | host: rabbit | reactivity:Arabidopsis thaliana

PRODUCT INFORMATION IN PDF

product information

background

Acd1 (accelerated cell death 1) EC=1.14.12.20 is an enzyme involved in chlorophyll breakdown, pheophorbide a oxygenase. This enzyme seems to induce cell death in Arabidopsis leaves in the dark. Synonymes:PaO, Lls1, lethal leaf-spot 1 homolog, pheide a oxygenase

immunogen

recombinant PaO from Arabidopsis thaliana Q9FYC2, At3g44880

antibody format

rabbit

polyclonal

serum

lyophilized

quantity

100 µl

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

western blot (WB)

related products

collection of antibodies to proteins involved in senescence

additional information

The protein level is moderately induced during dark-induced senescence.

application information
recommended dilution		1 : 5000 with standard ECL (WB)
expected \| apparent MW		61 \| 54 kDa
confirmed reactivity		Arabidopsis thaliana
predicted reactivity		dicots including: Brassica napus, Lycopersicum esculentum, Nicotiana tabacum
not reactive in		no confirmed exceptions from predicted reactivity are currently known
additional information		This antibody works on total cell extracts and can be used as a senescence marker. Predicted size of Acd1 precursor protein is about 61 kD including the transit peptide, but it must be processed to a smaller size. Using fresh extracts is recommended to decrease possible cross-reaction with Rubisco.
selected references		Nagane et al. (2010). Involvement of AtNAP1 in thre reulation of chlorophyll degradation in Arabiopsis thaliana. Planta (4):939-949. Hirashima et al. (2009). Light-independent cell death induced by accumulation of pheophorbide a in Arabidopsis thaliana. Plant Cell Physiol. (4):719-729.

application example

Arabidopsis thaliana wild ecotype Columbia was grown for four weeks under continuous illumination and then transferred to complete darkness for five days. Several leaves were harvested from the plants before they were transferred to darkness (0 d) or after they were kept for five days (5 d). Protein was extracted with the SDS extraction solution containing 50 mM Tris (pH 6.8), 10% (w/v) glycerol, 2% (w/v) SDS and 6% (v/v) 2-mercaptoethanol. Protein extract equivalent to 1 mg leaf material was loaded and separated on 14% SDS-PAGE and blotted 1h to PVDF. Blots were blocked with PBS-T containing 1.5% skim milk 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 PBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from GE Healthcare ) diluted to 1:20 000 in for 1h at RT with agitation. The blot was washed as above and developed for 1 min with ECLplus according to the manufacturers instructions. Exposure time was 5 min.

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