Anti-L32 | 50S ribosomal protein L32 (chloroplastic)

Product no: AS22 4864

AS22  4864 | Clonality: Polyclonal|  Host: Rabbit | Reactivity: Arabidopsis thaliana, Nicotiana tabacum

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  • Product Info
  • Immunogen: KLH-conjugated peptide derived from Arabidopsis thaliana L32 protein sequence, UniProt: P61847 and P12198 TAIR: ATCG01020
    Host: Rabbit
    Clonality: Polyclonal
    Purity: Antigern affinity purified serum, in PBS pH 7.4
    Format: Lyophilized
    Quantity: 50 µg
    Reconstitution: For reconstitution add 50 µl, of sterile or deionized 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: 6 kDa
  • Reactivity
  • Confirmed reactivity: Arabidopsis thaliana, Nicotiana tabacum
    Predicted reactivity: Brassica napus, Pisum sativum, Solanum lycopresicum, Solanum tuberosum

    Species of your interest not listed? Contact us
    Not reactive in: No confirmed exceptions from predicted reactivity are currently known
  • Application Examples
  • Western blot using anti-L32 (chloroplastic) antibodies

    1 - 10 ug of Nicotiana tabacum wild-type whole leaf extract
    2 - 10 ug of Nicotiana tabacum Rpl32 over-expresser whole leaf extract, contain native chloroplast Rpl32 (6.3 kDa) and fusion protein Rpl32-YFP (33.2 kDa)
    3 – 10 ug of Nicotiana tabacum wild-type root extract, used as negative control that does not contain chloroplast proteins
    ladder: MW markers

    Tobacco leaf and root protein was extracted freshly from Nicotiana tabacum (Exaction buffer components were: 0.7 M sucrose, 0.5 M Tris, 50 mM EDTA, 0.1 M KCL, 2% β-Mercaptoethanol, 2% Complete proteases inhibitor), mixed thoroughly and then added same volume of Phenol, precipitated in 0.1 M NH4OAc in Methanol, and resuspended in 10% SDS. Then denatured with 1 vol of 2x protein loading buffer (125 mM Tris-HCl (pH 6.8), 4% SDS, 20% glycerol, 25 mM EDTA, 0.4 mg/ml Bromophenol blue, 2% β-Mercaptoethanol at 64°C/10 min. 10 µg/well of protein samples were separated in the cold on 12% SDS-PAGE and blotted for 20 V/overnight to nitrocellulose (pore size of 0.2 um), using: wet transfer in the cold. Blot was blocked with 4 % milk for 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 for ON/4°C with agitation in TBS-T. The antibody solution was decanted, and the blot was rinsed briefly twice, then washed 3 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (Agrisera anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1: 10000 in TBS-T for 1h/RT with agitation. The blot was washed as above and developed with a following chemiluminescent detection reagents. Exposure time was 30 seconds.

    Courtesy of phd student. Jinghan Liu and Dr. Reimo Zoschke, Max Planck Institute, Germany

  • Background
  • Background: L32 (50S ribosomal protein L32) encodes a chloroplast ribosomal protein L32, a constituent of the large subunit of the ribosomal complex 
  • Protocols
  • Agrisera Western Blot protocol and video tutorials

    Protocols to work with plant and algal protein extracts

    Agrisera Educational Poster Collection
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