eIF2-alpha | Eukaryotic translation initiation factor 2 subunit alpha

AS20 4371 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

Product Information

Immunogen KLH-conjugated peptide derived from Arabidopsis thaliana eIF2-alpha, UniProt: Q9FE78, TAIR: AT5G05470

Host Rabbit

Clonality Polyclonal

Purity Affinity purified serum, in PBS pH 7.4

Format Lyophilized

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 Western blot (WB)

Recommended dilution 1 : 1000 (WB)

Expected | apparent MW 38.8 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana

Predicted reactivity Brassica napus, Brassica oleracea, Camelina sativa, Capsella rubella, Medicago truncatula, Noccaea caerulescens, Papaver somniferum, Raphanus sativus, Tarenaya hassleriana

Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples Western blot using anti-eIF2 antibodies

Samples:
1-4 14 days old Arabidopsis thaliana wild-type
5 - Arabidopsis thaliana gcn2 mutant

Arabidopsis thaliana whole seedlings were grinded in liquid nitrogen and 80 mg of tissue powder was resuspended in 100 µl of 0.5xSDS Laemmli buffer in a 1.5 ml tube. After 5 minutes of mixing at room temperature, tubes were centrifuged at 21 000 xg for 10 minutes at RT. For immunoblog analysis 10 µl of supernatant was separated on a 12.5 % (w/v) SDS-PAGE gel and electroblotted onto PVDF membrane. After 1 h of blocking at RT with 1x blocking buffer (TBS-T, 5 % non-fat milk and 0.2 % BSA), the membrane was washed with 1xTBS-T four times for 10 minutes each. Post washing, membrane was incubated with anti-eIF2alpha antibodies at 1: 5000 for 48 h at 4°C. Following the washing with TBS-T, 10 minutes each for six repeats, the membrane was incubated in the goat anti-rabbit IgG, HRP conjugated secondary antibodies for 1h at RT followed by washing with TBS-T 10 minutes each for ten repeats and the blot was developed with chemiluminescent detection reagents, according to manufacture's recommendations.

Courtesy of Dr. Ansul Lokdarshi, Biochemistry & Cellular and Molecular Biology, University of Tennessee, USA

Additional information

Related products

Related products Collection of antibodies involved in translation

Background

Background eIF2-alpha (Eukaryotic translation initiation factor 2 subunit alpha) functions in the early steps of protein synthesis by forming a ternary complex with GTP and initiator tRNA. eIF2 is a heterodimer composed of an alpha, beta and gamma chain.