Volume 2, Issue 1, Pages (January 2009)

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Volume 2, Issue 1, Pages 32-42 (January 2009) Immunoelectron Microscopy for Locating Calvin Cycle Enzymes in the Thylakoids of Synechocystis 6803 Rachna Agarwal, Stefan Ortleb, Jayashree Krishna Sainis, Michael Melzer Molecular Plant Volume 2, Issue 1, Pages 32-42 (January 2009) DOI: 10.1093/mp/ssn075 Copyright © 2009 The Authors. All rights reserved. Terms and Conditions

Figure 1 Electron Micrographs of Synechocystis Cells after High-Pressure Freezing. The log phase culture of Synechocystis 6803 was fixed through HPF and embedded in LR white. (A) Entire cells. (B) Portion of cell enlarged. Cx, carboxysome; T, thylakoid membrane. Space bars = 400 nm. Molecular Plant 2009 2, 32-42DOI: (10.1093/mp/ssn075) Copyright © 2009 The Authors. All rights reserved. Terms and Conditions

Figure 2 Immunogold Labeling of HPF Fixed Cells. Immunolabeling was carried out with polyclonal antibodies raised in rabbit. (A) anti-CF1. (B) anti-RPI. (C) anti-PRK. (D) anti-RuBisCO. (E) anti-PGK. (F) anti-GAPDH. 10 nm protein A-gold is shown for immunolabeling of CF1, RPI, RuBisCO, and PGK, 5 nm protein A-gold for PRK and GAPDH. Space bars = 200 nm. Molecular Plant 2009 2, 32-42DOI: (10.1093/mp/ssn075) Copyright © 2009 The Authors. All rights reserved. Terms and Conditions

Figure 3 Immunogold Labeling of the 150-k Thylakoid Membrane Fraction. Immunolabeling was carried out with polyclonal antibodies raised in rabbit. (A) anti-CF1. (B) anti-RPI. (C) anti-PRK. (D) anti-RuBisCO. (E) anti-GAPDH. (F) anti-PGK. Labeling done with 5 nm protein A-gold without primary antibody served as control. Arrows indicate the gold particles. Bar = 100 nm. Number of gold particles μm−2 were counted at 10 random locations. The average number of gold particles were 331.3 ± 38.8 for CF1; 325.6 ± 39.6 for RPI; 359.3 ± 18.2 for PRK; 354.9 ± 53.0 for RuBisCO; 366.2 ± 14.3 for PGK; and 282.9 ± 44.2 for GAPDH. The frequency of immunolabeling when no primary antibody was used was 95.7 ± 21.7 for protein A-gold particles of 5 nm in this experiment. Molecular Plant 2009 2, 32-42DOI: (10.1093/mp/ssn075) Copyright © 2009 The Authors. All rights reserved. Terms and Conditions