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CNX1/2 | CALNEXIN HOMOLOG 1/2

265 €
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AS12 2365 | clonality: polyclonal | host: rabbit | reactivity: Arabidopsis thaliana

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AS12 2365

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product information
Background

CNX1/2 (calnexin homolog 1/2) is a calcium-binding protein involved in protein folding. It interacts with newly synthesized glycoproteins in the endoplasmic reticulum.

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana  CNX1 UniProt: P29402 TAIR: AT5G61790, CNX2 UniProt: Q38798, TAIR: AT5G07340. This peptide is NOT present in calreticulins.

Host Rabbit
Clonality Polyclonal
Clone
Purity Affinity purified serum
Format Liquid
Quantity 50 µg
Reconstitution
Storage

Aliquite upon arrival to avoid repeated freeze-thaw cycles  and store  -20°C;  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
Western Blot (WB)
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AS10 683 | HDEL | endoplasmic reticulum retention signal (100 µg), mouse antibody
Additional information
application information
Recommended dilution

1 : 2500 with standard ECL (WB)

Expected | apparent MW
CNX1 60.5 kD, processing aa 1-20, mature peptide 58.1 kD

CNX2 60.5/61.4 kD, processing aa 1-25, mature peptides 57.6/58.6 kD
Confirmed reactivity Arabidopsis thaliana, Nicotiana tabacum
Predicted reactivity

Brassica napus, Medicago truncatula, Vitis vinifera, Oryza sativa, Solanum lycopersicum, Glycine max, Populus trichocarpa, Ricinus communis, Zea mays, Hordeum vulgare, Picea sitchensis, Pisum sativum, Physcomitrella patens, Coccomyxa suellipsoidea, Stereum hirsutum

Not reactive in

No confirmed exceptions from predicted reactivity are currently known

Additional information
Selected references Foley et al. (2017). A Global View of RNA-Protein Interactions Identifies Post-transcriptional Regulators of Root Hair Cell Fate.Dev Cell. 2017 Apr 24;41(2):204-220.e5. doi: 10.1016/j.devcel.2017.03.018.
LaMontagne et al. (2016). Isolation of Microsomal Membrane Proteins from Arabidopsis thaliana. Curr. Protoc. Plant Biol. 1:217-234. doi: 10.1002/cppb.20020.

Application example

western blot using anti-CNX1/2 antibodies

Total protein from Col-0 (wild-type) Arabidopsis thaliana were extracted with 50mM HEPES-KOH buffer containing 250 mM sucrose, 5% glycerol, 50 mM NaPP, 1 mM NaMo, 25 mM NaF, 10mM EDTA, 0.5% PVP, 3mM DTT, 1mM PMSF, 10uM Leupeptin & 10nM Calyculin, and then fractionated by ultracentrifugation at 100,000 x gravity for 30 min at 4°C into soluble (S100) and microsomal (P100) proteins as described in LaMontagne et al. (2016). Isolation of Microsomal Membrane Proteins from Arabidopsis thaliana. Current Protocols in Plant Biology 1:1-18. doi: 10.1002/cppb.20020. 30 µg proteins of total, S100 and P100 fractions were denatured at 37°C for 5 min, separated on a 7.5 % SDS-PAGE and blotted 1h to nitrocellulose using tank transfer. Blots were blocked with 1x PBS (from Fisher Scientific BP665-1) + 0.1 %Tween 20 (PBS-T) + 5% milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2500 overnight at 4°C with agitation in 1x PBS-T + 5% milk. The antibody solution was decanted, and the blot was rinsed briefly once, then washed four times for 7 min in 1x PBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in 1x PBS-T + 5% milk for 2 hrs at RT with agitation. The blot was washed as above and developed for 4 min with Amersham ECL (RPN2106). Exposure time was 30 seconds and 2 min

Courtesy of Erica LaMontagne & Dr. Antje Heese (Division of Biochemistry, Interdisciplinary Plant Group (IPG) - University of Missouri; Columbia, MO, USA)

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