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AKIN10 | SNF1-related protein kinase catalytic subunit alpha KIN10

AS10 919 Clonality: Polyclonal Host: Rabbit | Reactivity: Arabidopsis thaliana

AKIN10 | SNF1-related protein kinase catalytic subunit alpha KIN10 in the group Antibodies for Plant/Algal  / Developmental Biology / Signal transduction at Agrisera AB (Antibodies for research) (AS10 919)

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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

western blot using anti-AKIN10 antibodies


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.

Related products

Related products

AS09 460 | Anti-AKINB1 | SNF1-related protein kinase regulatory subunit beta-1, rabbit antibodies
AS09 462 | Anti-AKINB2 | SNF1-related protein kinase regulatory subunit beta-2, rabbit antibodies
AS15 2859 | Anti-AKIN ß3 | SNF1-related protein kinase regulatory subunit beta-3, rabbit antibodies
AS09 463 | Anti-AKIN beta gamma, rabbit antibodies
AS10 920 | Anti-AKIN11 | SNF1-related protein kinase catalytic subunit alpha KIN11, rabbit antibodies
AS09 613 | Anti-AKING1 | SNF1-related protein kinase regulatory subunit gamma 1, rabbit antibodies

collection of antibodies to proteins involved in signal transduction

Plant protein extraction buffer

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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