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MEB1 | Membrane protein of ER body 1

AS20 4423 | Clonality: Polyclonal  |  Host: Rabbit |  Reactivity: Arabidopsis thaliana

MEB1 | Membrane protein of ER body 1 in the group Antibodies for Plant/Algal  / Membrane Transport System / Endomembrane system at Agrisera AB (Antibodies for research) (AS20 4423)

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Product Information

Immunogen Purified recombinant MEB1 of Arabidopsis thaliana, residues 271-502 with a His tag, UniProt: Q8W4P8, TAIR: AT4G27860
Host Rabbit
Clonality Polyclonal
Purity Total IgG, purified on Protein A
Format Liquid at 2 mg/ml in PBS, 50% glycerol. Filter sterilized. No preservative or carrier added.
Quantity 100 µg
Storage Store at -20°C; once 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 material adhering to the cap or sides of the tubes.
Tested applications ELISA (ELISA), Immunoprecipitation (IP), Western blot (WB)
Recommended dilution assay dependent (ELISA), 1:100-1: 500 (IP), 1: 1000-1: 2000 (WB)
Expected | apparent MW 68 | 85 kDa (due to a large number of hydrophobic residues)

Reactivity

Confirmed reactivity Arabidopsis thaliana
Predicted reactivity Species of your interest not listed? Contact us
Not reactive in No confirmed exceptions from predicted reactivity are currently known.

Application examples

Application examples Western blot using anti-MEB1 antibodies

Arabidopsis thaliana 7 day-old seedlings were freshly extracted with 2x SDS-sample buffer (+ 2ME) for SDS-PAGE and denatured with 4X SDS buffer at 95°C for 5 min. Protein load/well is 10 µg. Sample was separated on 12.5 % SDS-PAGE and blotted at 15V overnight using wet transfer to PVDF membrane. Blot was blocked with 3 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2000 in TBS-T for 1h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendation.
Calculated MW of MEB1 is 68 kDa, while apparent MW appears to be 85 kDa (due to a large number of hydrophobic residues)


Western blot using anti-MEB1 antibodies

Samples of 7-day old seedlings from Arabidopsis thaliana wild-type (1), mutant meb1-1 (2), mutant meb1-2 (3), mutant meb1-3 (4), mutant meb2-1 (5), mutant meb2-3 (6), mutant meb1-1 meb2-1 (7), mutant nal-1-1 (8) were freshly extracted with 2x SDS-sample buffer (+ 2ME) for SDS-PAGE and denatured with 4X SDS buffer at 95°C for 5 min. Samples were separated on 10% SDS-PAGE and blotted 1h to nitrocellulose membrane. Blot was blocked with 3 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2000 in TBS-T for 1h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendation. Coomassie blue staining (CBB) shows the Rubisco large subunit, which served as aloading control.
NAI1 protein is MEB1 interacting protein. 

Additional information

Additional information This antibody does not detect MEB2 protein in Arabidopsis thaliana.

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Background

Background MEB1 (Membrane protein of ER body 1) displays iron ion transmembrane transporter activity and may sequester excess cytosolic iron and manganese into endoplasmic reticulum to reduce metal ion toxicity. Not essential for the accumulation of ER body components.

Product citations

Selected references Yamada et al. (2013). Identification of two novel endoplasmic reticulum body-specific integral membrane proteins. Plant Physiol . 2013 Jan;161(1):108-20. doi: 10.1104/pp.112.207654.

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