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RCD1 | Radical cell death 1
Product no: AS12 2602
AS12 2602 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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319
€
Delivery:
3-6 business days
- Data sheet
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- Product Info
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Immunogen: KLH-conjugated synthetic peptide derived from Arabidopsis thaliana RCD1 protein sequence UniProt: Q8RY59, TAIR: AT1G32230
Host: Rabbit Clonality: Polyclonal Purity: Immunogen affinity purified serum in PBS pH 7.4. Format: Lyophilized Quantity: 100 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 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) following immunoprecipitation (IP) Recommended dilution: 1 : 1000 (WB) on samples following IP Expected | apparent MW: 65 kDa
- Reactivity
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Confirmed reactivity: Arabidopsis thaliana Not reactive in: No confirmed exceptions from predicted reactivity are currently known - Application Examples
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application information
5 mg of total protein from Arabidopsis thaliana seedlings expressing RCD1-HA under native promoter extracted with RIPA buffer for protein extraction (50 mM Tris-HCl pH 7,5, 150 mM NaCl, 1% Triton X-100, 0.5% sodium deoxycholate, protease inhibitor cocktail 1:100, 50 uM MG132) were used for immunoprecipitation and afterwards separated on 10% SDS-PAGE and blotted 1h to PVDF using tank transfer. Blots were blocked with 5% milk for 1h at room temperature (RT) with agitation. Blot was incubated in the anti-HA antibody (Roche) at a dilution of 1: 1 000 in 1% milk, TBS-T overnight at +4C 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-rat IgG horse radish peroxidase conjugated) diluted to 1:10 000 in 1% milk TBS-T for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL Prime, GE Healthcare. Exposure time was 1 min.
Courtesy Dr. Julia Vainonen, Helsinki University, Finland
5 µl of anti-RCD1 antibody was used for immunoprecipiation. Samples: transgenic line expressing HA-tagged RCD1 (Jaspers et al., 2009, Plant J) (1), rcd1-4 mutant (2). Afterwards these samples were separated on 10% SDS-PAGE and blotted 1h to PVDF using tank transfer. Blots were blocked with 5% milk for 1h at room temperature (RT) with agitation. Blot was incubated in the anti-HA antibody (Roche) at a dilution of 1: 1 000 in 1% milk, TBS-T overnight at +4C 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-rat IgG horse radish peroxidase conjugated) diluted to 1:10 000 in 1% milk TBS-T for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL Prime, GE Healthcare. Exposure time was 1 min.
Courtesy Dr. Julia Vainonen, Helsinki University, Finland - Additional Information
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Additional information (application): Protein has three splice variants: 63.8 + 65.6 + 65.7 kD
Peptide used to elicit this antibody was designed for detection of all (3) RCD1 splice variants but NOT the close relative SRO1.
- Background
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Background: RCD1 (Radical cell death 1) functions as a regulator of oxidative-stress hormonal and developmental responses. Alternative name: Inactive poly [ADP-ribose] polymerase RCD1. - Protocols
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