AKIN ß gamma
AS09 463 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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Product Information
Immunogen
KLH-conjugated peptide derived from Arabidopsis thaliana AKIN beta gamma, UniProt: Q944A6, TAIR: AT1G09020
Host
Rabbit
Clonality
Polyclonal
Purity
Affinity purified serum in PBS pH 7.4
Format
Lyophilized
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)
Expected | apparent MW
53 kDa
Reactivity
Confirmed reactivity
Arabidopsis thaliana
Application examples
Application examples
Application example
#1: 5μg of purified (Ni-NTA, Qiagen, manufacturer’s instruction) recombinant protein expressed in E. coli were loaded.
#2: Protoplast sample, AKINbg-HA was overexpressed as in Ramon et al (2013, the Plant Journal) and boiled in Laemmli solubilisation buffer for 5min. The proteins were separated on 8% SDS-PAGE and blotted 1h to PVDF using semi-dry transfer. Blots were blocked with 5% non-fat Milk in TBS for 1h at room temperature (RT) with agitation. Blots were incubated in the primary antibody at a dilution of 1:1000 for 12h at 4°C with agitation. The antibody solution was removed and the blot was then washed four times for 5 min in TBS-T at RT with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in 1% non-fat milk-TBS for 1h at RT with agitation. The blots were washed as above, 5 times, and developed for 5 min with ECL according to the manufacturer's instructions. Exposure time was 20 seconds for the recombinant proteins (#1) or 5 min for protoplasts sample (#2).
Courtesy of Dr Pierre Crozet and Dr Elena Baena-Gonzalez. Spain
#1: 5μg of purified (Ni-NTA, Qiagen, manufacturer’s instruction) recombinant protein expressed in E. coli were loaded.
#2: Protoplast sample, AKINbg-HA was overexpressed as in Ramon et al (2013, the Plant Journal) and boiled in Laemmli solubilisation buffer for 5min. The proteins were separated on 8% SDS-PAGE and blotted 1h to PVDF using semi-dry transfer. Blots were blocked with 5% non-fat Milk in TBS for 1h at room temperature (RT) with agitation. Blots were incubated in the primary antibody at a dilution of 1:1000 for 12h at 4°C with agitation. The antibody solution was removed and the blot was then washed four times for 5 min in TBS-T at RT with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in 1% non-fat milk-TBS for 1h at RT with agitation. The blots were washed as above, 5 times, and developed for 5 min with ECL according to the manufacturer's instructions. Exposure time was 20 seconds for the recombinant proteins (#1) or 5 min for protoplasts sample (#2).
Courtesy of Dr Pierre Crozet and Dr Elena Baena-Gonzalez. Spain
Additional information
This antibody is recognizing well recombinant AKIN beta gamma as well as the over-expressed protein in protoplasts.
It has not been confirmed working on endogenous cell extract.
It has not been confirmed working on endogenous cell extract.
Background
Background
AKIN beta gamma is a regulatory subunit of SNF1-related protein kinase (SnRK) complex. Involved in signal transduction cascade in carbohydrate metabolism. Alternative names: ATSNF4, homolog of yeast sucrose nonfermenting 4, KIN??, SNF4.
Product citations
Selected references
Emanuelle et al. (2015). SnRK1 from Arabidopsis thaliana is an atypical AMPK. Plant J. 2015 Mar 3. doi: 10.1111/tpj.12813.
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