BAK1 | Brassinosteroid insensitive 1-associated receptor kinase 1

AS12 1858 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Solanum lycopersicum

BAK1 | Brassinosteroid insensitive 1-associated receptor kinase 1 in the group Plant/Algal Antibodies / Hormones / Brassinosteroids at Agrisera AB (Antibodies for research) (AS12 1858)


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BAK1 (Brassinosteroid insensitive 1-associated receptor kinase 1) controls expression of genes associated with innate immunity in the absence of pathogens or elicitors. Phosphorylated Brl1. Involved in programmed cell death (PCD). Subcellular localization: cell membrane. Alternative names: BRASSINOSTEROID INSENSITIVE 1-associated receptor kinase 1, BAK1, BRI1-ASSOCIATED RECEPTOR KINASE, RKS10, RECEPTOR KINASES LIKE SERK 10, SERK3, SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE 3, ELG, ELONGATED, ATSERK3, SOMATIC EMBRYOGENESIS RECEPTOR-LIKE KINASE 3, ATBAK1.

Immunogen KLH-conjugated peptide, chosen from Arabidopsis thaliana Bak1 sequence, TAIR: AT4G33430 UniProt: Q94F62
Host Rabbit
Clonality Polyclonal
Purity Affinity purified serum in PBS, pH 7.4
Format Lyophilized in PBS pH 7.4.
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 Immunoprecipitation (IP), Western blot (WB)
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AS16 3204 | Anti-SOBIR1 | supressor of BIR1 antibody

Plant protein extraction buffer

Secondary antibodies

Additional information
application information
Recommended dilution 2 µl/50 µl of Protein G agarose, 1 : 5000 (WB)
Expected | apparent MW

68 | 70 kDa

Confirmed reactivity Arabidopsis thaliana, Solanum lycopersicum
Predicted reactivity Thelungiella halophila
Not reactive in

Hordeum vulgare, Nicotiana benthamiana, Oryza sativa

Additional information Extra Information on CE extraction buffer: CE buffer does not need to be made freshly everytime. Aliquots can be kept at -20°C. Na2MoO4 and NaF are phosphatase inhibitors, included to prevent lose phosphorylation form our protein of interest during extraction. EDTA chelates metal ions and thus inhibits many enzymes which need metal ions as co-factors and inhibits the action of proteases.  Protease inhibitor coctail is Sigma product number P 9599 which is used in dilution 1: 100.
Selected references Hu et al. (2018). A group of receptor kinases are essential for CLAVATA signalling to maintain stem cell homeostasis. Nat Plants. 2018 Apr;4(4):205-211. doi: 10.1038/s41477-018-0123-z. (CoIP)
Bundy et al. (2016). A mutation in the catalytic subunit of the glycosylphosphatidyl inositol transamidase disrupts growth, fertility and stomata forma formation in Arabidopsis. Plant Physiology. DOI:10.1104/pp.16.00339.
Tateda et al. (2014). Salicylic Acid Regulates Arabidopsis Microbial Pattern Receptor Kinase Levels and Signaling.Plant Cell. 2014 Oct 14. pii: tpc.114.131938.

Application information

western blot using anti-Bak1 antibodies

Plant material: Arabidopsis thaliana Col-0 (wild type) bak1-4 (knock-out mutant for BAK1, does not show any full-length BAK1 transcript, Kemmerling et al., 2007). Method: Two-week-old seedlings and leaf material of 5-week-old plants (grown under short day conditions, 8h light) were collected and frozen in liquid nitrogen. Approximately 100 mg of plant material were extracted in 0.2 ml of homogenization buffer (250 mM sucrose, 50 mM HEPES-KOH pH 7.5, 0.5% Triton X-100, 5% glycerol, 50 mM Na4P2O7, 1 mM Na2MoO4, 25 mM NaF, 2 mM DTT, Sigma plant protease inhibitor cocktail) with a glass pistil and a small amount of sand in an 1.5ml Eppendorf tube. Another 0.8 ml of buffer were added and the extract was mixed thoroughly. Debris was pelleted by centrifugation in a table-top microcentrifuge at 13 000 rpm. The supernatant was mixed with 4x SDS loading buffer (200 mM TRIS-HCl pH 6.8, 400 mM DTT, 8% SDS, 40% glycerol, 0.1% bromophenol blue) and boiled at 95°C for 5min. 10µl were run on an 10% polyacrylamide gel and blotted onto a 0.45µm PVDF membrane (Carl Roth). The membrane was blocked for 1h in TBS-T (150mM NaCl, 10mM Tris-HCl pH8, 0.05% Tween-20) containing 5% skimmed milk powder. The primary antibody (Agrisera Rabbit anti-BAK1, AS12 1858, 1µg/µl) was diluted 1:5000 in TBS-T containing 5% milk powder and incubated on the membrane overnight at 4°C. Then the membrane was washed 5 times 15min with TBS-T containing 5% milk powder. The secondary antibody (Agrisera Goat anti-rabbit IgG (H&L) HRP conjugate, AS09 602, 0.91 µg/µl) was diluted 1:5000 in the same solution and incubated on the membrane at room temperature for 2h. The membrane was then washed 5 times 15min with TBS-T (no milk powder) and the blot was developed using Super Signal West Pico Chemiluminescent Substrate (Thermo Scientific). Short exposure: 1 minute. Long exposure: 10 minutes.

Courtesy of Dr. Elena Petusching, Georg-August-University Goettingen, Germany

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