AKIN10 | SNF1-related protein kinase catalytic subunit alpha KIN10
AS10 919 Clonality: Polyclonal Host: Rabbit | Reactivity: Arabidopsis thaliana
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Product Information
Immunogen
KLH-conjugated synthetic peptide derived from C-terminal part of Arabidopsis thaliana AKIN10 sequence UniProt: Q38997, TAIR: At3g01090
Host
Rabbit
Clonality
Polyclonal
Purity
Affinity purified serum in PBS, pH 7.4
Format
Lyophilized
Quantity
100 µg
Reconstitution
For reconstitution add 50 µl/tube 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
Immunolocalization (IL), Western blot (WB)
Recommended dilution
1: 50 (IL), 1 : 500 (WB)
Expected | apparent MW
61.1 kDa
Reactivity
Confirmed reactivity
Arabidopsis thaliana
Predicted reactivity
Arabidopsis thaliana
Not reactive in
Oryza sativa, Solanum lycopersicum, Vitis vinifera
Application examples
Application examples
Application example
Arabidopsis thaliana total proteins were extracted in Hepes buffer (50mM Hepes-NaOH pH 7.8; 2mM EDTA pH 8.0; 1mM DTT; Phosphatase and Protease inhibitor cocktails). After centrifugation, the supernatant was recovered and protein concentration determined using the Bradford protein assay. 5, 10, 20 and 30 μg of total protein were resolved, for each plant extract, by SDS-PAGE, transferred to a PVDF membrane and analyzed by immunoblotting with α-AKIN10 antibody (Agrisera; AS10 919; 1:500; 1% non-fat Milk in TBS O/N at 4°C). Secondary antibody anti-rabbit HRP-conjugated was used at 1:20 000, 1% non-fat Milk in TBS for 2h at RT. Washes were made as following: after blocking, the membrane was washed 2X 5 min with 1x TTBS; after the primary antibody incubation, the membrane was washed 5x 5 min with 1x TTBS; after the secondary antibody incubation, the membrane was washed 5x 5 min with 1x TTBS (contained Tween-20 at 500 µl/liter), followed by 3x 5 min with 1x TBS Wash Buffers (TBS and TTBS): 1X TBS (1 liter) 2.42 g Tris 8 g NaCl Adjust pH to 7.5-7.6 with HCl 1X TTBS: 1X TBS + 500 ul Tween-20 per liter. Reaction was developed following manufacture's recommendations and recorded using BioRad GelDoc.
Courtesy of Drs. Leonor Margalha/Elena Baena-González, Instituto Gulbenkian De Ciencia, Portugal
Arabidopsis thaliana total proteins were extracted in Hepes buffer (50mM Hepes-NaOH pH 7.8; 2mM EDTA pH 8.0; 1mM DTT; Phosphatase and Protease inhibitor cocktails). After centrifugation, the supernatant was recovered and protein concentration determined using the Bradford protein assay. 5, 10, 20 and 30 μg of total protein were resolved, for each plant extract, by SDS-PAGE, transferred to a PVDF membrane and analyzed by immunoblotting with α-AKIN10 antibody (Agrisera; AS10 919; 1:500; 1% non-fat Milk in TBS O/N at 4°C). Secondary antibody anti-rabbit HRP-conjugated was used at 1:20 000, 1% non-fat Milk in TBS for 2h at RT. Washes were made as following: after blocking, the membrane was washed 2X 5 min with 1x TTBS; after the primary antibody incubation, the membrane was washed 5x 5 min with 1x TTBS; after the secondary antibody incubation, the membrane was washed 5x 5 min with 1x TTBS (contained Tween-20 at 500 µl/liter), followed by 3x 5 min with 1x TBS Wash Buffers (TBS and TTBS): 1X TBS (1 liter) 2.42 g Tris 8 g NaCl Adjust pH to 7.5-7.6 with HCl 1X TTBS: 1X TBS + 500 ul Tween-20 per liter. Reaction was developed following manufacture's recommendations and recorded using BioRad GelDoc.
Courtesy of Drs. Leonor Margalha/Elena Baena-González, Instituto Gulbenkian De Ciencia, Portugal
Additional information
AKIN10 antibody works best when first the SnRK1 complex is immunorecipitated with this or another antibody and then anti-AKIN10 antibody is used to probe with. When this antibody is used against a lysate, the dominant rubisco protein can mask any immunoreactivity.
Background
Background
AKIN10 (E.C.= 2.7.11.1) is a catalytic subunit of the putative trimeric SNF1-related protein kinase (SnRK) complex, which may play a role in a signal transduction cascade regulating gene expression and carbohydrate metabolism in higher plants. Synonymes: AKIN alpha-2
Product citations
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