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AKIN ß gamma

AS09 463 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

AKIN ß gamma in the group Antibodies Plant/Algal  / Plant Developmental Biology / Plant Signal Transduction at Agrisera AB (Antibodies for research) (AS09 463)
AKIN ß gamma



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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 Immunogen affinity purified serum in PBS pH 7.4.
Format Lyophilized
Quantity 50 ĩg
Reconstitution For reconstitution add 50 ĩl 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)
Expected | apparent MW

53 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana

Application examples

Application examples Application example
western blot using anti-AKIN beta gamma antibodies
#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.

Related products

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.
immunogen: KLH-conjugated peptide derived from Arabidopsis thaliana AKIN beta gamma, UniProt: Q944A6, TAIR: AT1G09020
Reconstitution: For reconstitution add 50 ĩl of sterile water
Host: Rabbit
Clonality: Polyclonal
Purity: Immunogen affinity purified serum in PBS pH 7.4.
Format: Lyophilized
Quantity: 50 ĩg
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)
calculated | apparent molecular mass [kDa]:

53 kDa

Confirmed reactivity: Arabidopsis thaliana
Picture (footer): Application example
western blot using anti-AKIN beta gamma antibodies
#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 (application): 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.
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.
All 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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