Total plant protein from the greenhouse grown young leaves or the powdered lyophilized plant cells of transplastomic lettuce expressing CTB-VP1n and CTB-VP1co, and wild-type were extracted in the protein extraction buffer (100 mM NaCl, 10 mM EDTA, 200 mM Tris-Cl pH 8.0, 0.05% (v/v) Tween-20, 0.1% SDS, 14 mM β-ME, 400 mM sucrose, 2 mM PMSF, and proteinase inhibitor cocktail). Total protein extracts were analyzed by 10% SDS-PAGE. Western blot analysis was performed by rabbit anti-CTB polyclonal primary antibody (GenWay Biotech Inc, San Diego, CA) in a dilution of 1:10,000 followed by detection with a goat anti-rabbit IgG-HRP secondary antibody (1: 4,000 dilution; SouthernBiotech, Birmingham, AL) and developed using the ECL chemiluminescence (GE healthcare, UK). CTB (Cat# GWB-7B96E4, GenWay Biotech Inc, San Diego, CA) was used as positive control and served as standard for quantitative analysis of the blot with Image J software (IJ 1.46r; NIH). For loading controls, protein-blotted membrane was stained with Ponceau S (Sigma, P-3504) prior to immunoprobing with anti-CTB antibody and anti-RbcL antibody (Cat# AS03 037, Agrisera, 1:5,000) was used.
Anti rbcl antibody
Manufactured by Agrisera
The Anti-RbcL antibody is a laboratory tool used to detect and quantify the RbcL protein, a critical component of the Rubisco enzyme involved in photosynthesis. This antibody can be utilized in various analytical techniques, such as Western blotting, to study the expression and localization of the RbcL protein in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Ecl chemiluminescence, by GE Healthcare (2 mentions)
Goat anti rabbit igg hrp secondary antibody, by Southern Biotech (2 mentions)
Rabbit anti ctb polyclonal primary antibody, by Aviva Systems Biology (2 mentions)
Ponceau s, by Merck Group (1 mentions)
Glutathione resin, by GenScript (1 mentions)
Rproteina beads 4ff, by Smart-Lifesciences (1 mentions)
Alexa fluor 488 labeled goat anti mouse igg antibodies, by Beyotime (1 mentions)
Ni nta beads 6ff, by Smart-Lifesciences (1 mentions)
Anti gfp affinity beads, by Smart-Lifesciences (1 mentions)
Nitrocellulose membrane, by GE Healthcare (1 mentions)
4 protocols using anti rbcl antibody
1
Quantifying Transplastomic Lettuce Protein
2
Quantitative Analysis of Transplastomic Lettuce Protein
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Total plant protein from the greenhouse grown young leaves or the powdered lyophilized plant cells of transplastomic lettuce expressing CTB‐VP1n and CTB‐VP1co, and wild‐type were extracted in the protein extraction buffer (100 mm NaCl, 10 mm EDTA, 200 mm Tris‐Cl pH 8.0, 0.05% (v/v) Tween‐20, 0.1% SDS, 14 mm β‐ME, 400 mm sucrose, 2 mm PMSF and proteinase inhibitor cocktail). Total protein extracts were analysed by 10% SDS‐PAGE. Western blot analysis was performed by rabbit anti‐CTB polyclonal primary antibody (GenWay Biotech Inc, San Diego, CA) in a dilution of 1:10 000 followed by detection with a goat anti‐rabbit IgG‐HRP secondary antibody (1: 4000 dilution; SouthernBiotech, Birmingham, AL) and developed using the ECL chemiluminescence (GE healthcare, Stevenage, UK). CTB (Cat# GWB‐7B96E4; GenWay Biotech Inc, San Diego, CA) was used as positive control and served as standard for quantitative analysis of the blot with Image J software (IJ 1.46r; NIH). For loading controls, protein‐blotted membrane was stained with Ponceau S (Sigma, P‐3504) prior to immunoprobing with anti‐CTB antibody and anti‐RbcL antibody (Cat# AS03 037, Agrisera, 1:5000) was used.
3
Antibody and Affinity Reagents for Protein Detection
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The following antibodies were purchased: anti-Myc antibody (GNI, GNI4410-MC, 1:4000); anti-Flag antibody (GNI, GNI4410-FG, 1:4000); anti-H + -ATPase antibody (PHYTOAB, PHY2285A, 1:2000); anti-Rbcl antibody (Agrisera, AS03037, 1:4000); anti-H3 antibody (Abmart, P30266M, 1:3000); anti-β-Tubulin antibody (Abmart, M30109M, 1:4000); anti-GFP antibody (Abmart, M20004, 1:3000); anti-PIF4 antibody (Abiocode, R2534-4, 1:2000); anti-GST antibody (Abmart, M20007, 1:4000); anti-His antibody (GNI, GNI4110-HS, 1:4000); Alexa Fluor 488-labeled Goat Anti-Mouse IgG antibodies (Beyotime, A0428, 1:200); Glutathione Resin (GenScript, L00206); anti-GFP Affinity beads (Smart-Lifesciences, SA070001); rProtein A Beads 4FF (Smart-Lifesciences, SA015005); Ni NTA Beads 6FF (Smart-Lifesciences, SA005025).
4
Immunoblotting for RbcL Protein
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Immunoblotting was carried out by resolving secreted and calli proteins on 12.5% SDS-PAGE followed by electrotransfer onto nitrocellulose membrane (GE Healthcare). The blot was probed with anti-RbcL antibody (AS03037; Agrisera AB) at a dilution of 1:5000 in TBS. Immunoreactive protein was detected by incubation with alkaline phosphatase conjugated anti-rabbit IgG as secondary antibody (Sigma).
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