COI1 | Coronate insensitive 1 (rabbit antibody)
AS12 2637 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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Product Information
Immunogen
KLH-conjugated synthetic peptide derived from Arabidopsis thaliana COI1 protein, UniProt: O04197 , TAIR: At2g39940
Host
Rabbit
Clonality
Polyclonal
Purity
Immunogen affinity purified serum in PBS pH 7.4.
Format
Lyophilized
Quantity
50 µg
Reconstitution
For reconstitution add 25 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 the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Tested applications
Western blot (WB)
Recommended dilution
1 : 1000 (WB)
Expected | apparent MW
70 kDa
Reactivity
Confirmed reactivity
Arabidopsis thaliana
Predicted reactivity
Brassica rapa, Glycine max, Lupinus luteus, Medicago tribuloides, Nicotiana tabacum, Oryza sativa, Pisum sativum, Populus trichocarpa, Ricinus communis, Solanum lycopersicum, Vitis vinifera
Species of your interest not listed? Contact us
Species of your interest not listed? Contact us
Not reactive in
No confirmed exceptions from predicted reactivity are currently known
Application examples
Application examples
Total proteins were isolated from 7-day old Arabidopsis thaliana Col-0 (wild-type) and coi1 (null mutant) seedlings according to [ref 1]. Proteins were denatured at 65°C for 5 min. 30 μg of proteins were separated on a 10% SDS-PAGE and transferred for 70 min at 55V using a tank transfer system to nitrocellulose membrane. Blots were blocked with 1x PBS + 0.1% Tween 20 (PBS- T) + 5% milk for 1hr at room temperature (RT) with agitation. Blots were incubated in primary ⍺-COI1 antibody (AS12 2637) diluted to 1:5,000 in PBS-T+5 % milk at 4°C overnight with agitation. The primary antibody solution was decanted, and the blots were washed four times (6-8 minutes each) in 1x PBS-T at RT with agitation prior to incubation with secondary antibody Goat anti Rabbit IgG (H&L) – HRP conjugated (AS09 602-trial) diluted to 1:20,000 in 1x PBS-T + 5% milk for 2hr at RT with agitation. The blots were washed as above and developed for 4 min with chemiluminescent detection reagent ECL Bright (AS16 ECL-N). Exposure times (in seconds) to X-ray films were done as indicated. PonceauS stained membrane and membrane portion probed with ⍺-MPK6 served as loading controls [ref 1].
Courtesy of Alani Antoine-Mitchell and Antje Heese, University of Missouri, Div. Biochemistry, IPG (USA)
References: [1] LaMontagne, ED., Collins, CA., Peck, S.C., and Heese, A. 2016. Isolation of microsomal membrane proteins from Arabidopsis thaliana. Curr. Protoc. Plant Biol. 1:217-234. doi: 10.1002/cppb.20020
Total proteins were isolated from 7-day old Arabidopsis thaliana Col-0 (wild-type) and coi1 (null mutant) seedlings according to [ref 1]. Proteins were denatured at 65°C for 5 min. 30 μg of proteins were separated on a 10% SDS-PAGE and transferred for 70 min at 55V using a tank transfer system to nitrocellulose membrane. Blots were blocked with 1x PBS + 0.1% Tween 20 (PBS- T) + 5% milk for 1hr at room temperature (RT) with agitation. Blots were incubated in primary ⍺-COI1 antibody (AS12 2637) diluted to 1:5,000 in PBS-T+5 % milk at 4°C overnight with agitation. The primary antibody solution was decanted, and the blots were washed four times (6-8 minutes each) in 1x PBS-T at RT with agitation prior to incubation with secondary antibody Goat anti Rabbit IgG (H&L) – HRP conjugated (AS09 602-trial) diluted to 1:20,000 in 1x PBS-T + 5% milk for 2hr at RT with agitation. The blots were washed as above and developed for 4 min with chemiluminescent detection reagent ECL Bright (AS16 ECL-N). Exposure times (in seconds) to X-ray films were done as indicated. PonceauS stained membrane and membrane portion probed with ⍺-MPK6 served as loading controls [ref 1].
Courtesy of Alani Antoine-Mitchell and Antje Heese, University of Missouri, Div. Biochemistry, IPG (USA)
References: [1] LaMontagne, ED., Collins, CA., Peck, S.C., and Heese, A. 2016. Isolation of microsomal membrane proteins from Arabidopsis thaliana. Curr. Protoc. Plant Biol. 1:217-234. doi: 10.1002/cppb.20020
Additional information
Antibody detects 100 ng of recombinant COI1 protein, expressed in Hi5 suspenstion insect cells.
The peptide used to elicit this antibody is partially conserved in Zea mays: ZmCOI1a, ZMCOI1b, ZMCO1c and ZmCOI2.
Background
Background
COI1 (Coronate insensitive 1) is an esential component of jasmonate receptor. Required for jasmonate-regulated plant fertility and defense processes, and for coronatine and/or other elicitors perceptions/responses. Alternative names: F-box/LRR-repeat protein 2, AtCOI1, AtFBL2.
Product citations
Selected references
Agrawal et al. (2022) MEDIATOR SUBUNIT17 integrates jasmonate and auxin signaling pathways to regulate thermomorphogenesis. Plant Physiol. 2022 Aug 1;189(4):2259-2280. doi: 10.1093/plphys/kiac220. PMID: 35567489.
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