Anti-Toc75 | Protein TOC75-3, chloroplastic, POTRA domain 1
Referentie AS08345
Formaat : 200ul
Merk : Agrisera
Anti-Toc75 | Protein TOC75-3, chloroplastic, POTRA domain 1
AS08 345 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Pisum sativum
- YI-SHAN TENG | 2012-05-03I used this antibody for western blotting with 1/1000 dilution on 20-30 ug of chloroplast protein/lane with coloring method. The titer of this antibody is very good and it can recognize Toc75 very well in pea chloroplasts.
| Immunogen: | psTOC75; Predicted POTRA Domain #1; Amino acids, 158-241; Expressed and purified in E. coli using the Impact System from NEB. Peptide confirmed by MALDI. Q43715 |
| Host: | Rabbit |
| Clonality: | Polyclonal |
| Purity: | Serum |
| Format: | Lyophilized |
| Quantity: | 200 µl |
| Reconstitution: | For reconstitution add 200 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 the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube. |
| Tested applications: | Flow cytometry (Flow cyt), Immunolocalization (IL),Western blot (WB) |
| Recommended dilution: | 1 : 500 (Flow cyt), 1 : 100 (IL), 1 : 2000-1 : 100 000 (WB) |
| Expected | apparent MW: | 88 | 75 kDa (ocassionally a processing intermediate at 78 kDa is observed) |
| Confirmed reactivity: | Pisum sativum, some cross-reactivity was observed for cyanobacteria including: Synechocystis, Synechococcus and Thermosynechococcus |
| Predicted reactivity: | Oryza sativa, Ricinus communis, Populus trichocarpa, Vitis vinifera Species of your interest not listed? Contact us |
| Not reactive in: | Physcomitrella patens, Zea mays |
| Application example
580 ng of Chl of Pisum sativum plants (10 day old) (2, 4) and 10 µg of combined envelopes of Pisum sativum 10 day-old (1,3) were separated on 15% SDS-PAGE and blotted 2h to PVDF. PVDF was blocked 1h with 3% non-fat milk powder in TBS-T (0.1% TWEEN 20) and probed with anti-Toc75 POTRA domain 1 antibodies AS08 345 (1:2000 and 1: 5000, 1h) and secondary donkey-anti-rabbit (1:20000, 1 h) antibody (HRP conjugated) in TBS-T containing 3% non fat milk powder. Antibody incubations were followed by washings in TBS-T. All steps were performed at RT with agitation. Blots were developed for 5 min with HRP substrate Peroxide solution & luminol detection reagent according to the manufacturers instructions. Exposure time was 600 seconds. Courtesy of Ashita Dave and Barry D. Bruce (UTK), USA |
| Additional information (application): | Antibody detects Toc75 POTRA domain 1 as purified protein, in chloroplast fraction and in crude envelope fraction |
| Background: | OEP 75 or Toc75; Chloroplast outer envelope membrane protein from Pisum sativum (pea), Predicted to contain 3 POTRA domains at N-terminus. Believed to be the protein conducting channel of the Toc translocon and assembles as an 18 stranded ß-barrel (EMBO J. (1995) 14:11, 2436-2446); In Arabidopsis there are five members of this Family Toc75 (I-V), atToc75III is most closely related to psToc75. Additionally, it is structurally related to members of the bacterial surface antigen super-family including: OMA87; Outer membrane protein/protective antigen, (COG4775, COG4775) [Cell envelope biogenesis, outer membrane; YaeT; outer membrane protein assembly complex, (TIGR03303); FhaC; Hemolysin activation/secretion protein (COG2831) [Intracellular trafficking and secretion] |
Protocols to work with plant and algal protein extracts
Oxygenic photosynthesis poster by prof. Govindjee and Dr. Shevela
Z-scheme of photosynthetic electron transport by prof. Govindjee and Dr. Björn and Dr. Shevela