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Anti-CAH3 | Carbonic anhydrase
AS05 073 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Chlamydomonas reinhardtii
- Product Info
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Immunogen: Recombinant carbonic anhydrase of Chlamydomonas reinhardtii A8J4Z8 after cleavage of a fusion protein Host: Rabbit Clonality: Polyclonal Purity: Serum Format: Lyophilized Quantity: 50 µl Reconstitution: For reconstitution add 100 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: immunofluorescence (IF), Immunogold (IG), Western blot (WB) Recommended dilution: 1 : 200 (IF), 1 : 2000 (WB) Expected | apparent MW: 33.4 | 29 kDa - Reactivity
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Confirmed reactivity: Chlamydomonas reinhardtii Predicted reactivity: Chlamydomonas reinhardtii (only)
Not reactive in: Arabidopsis thaliana, Oryza sativa
- Application Examples
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Application example
(1) Chlamydomonas reinhardtii thylakoid preparation, (2) overexoressed carbonic anhydrase, primary antibody used in 1: 2000 dilution. Secondary antibodies anti-rabbit IgG. used at 1: 10 000 with chemiluminescent detection.
20 µg/ml of chlorophyll from Chlamydomonas reinhardtii Cah3-less mutant (1), and 15 µg/ml of chlorophyll from Chlamydomonas reinhardtii wild-type (2),extracted with 4x Laemmli were separated on 12 % SDS-PAGE and blotted 2h to nitrocellulose filter using tank transfer. Blots were blocked with 5% milk powder for for 2h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 fover night at 8°C with agitation. The antibody solution was decanted and the blot was rinsed briefly, then washed once for 15 min and 3 times for 20 min each in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:20 000 in for 1h at RT with agitation. The blot was washed as above and developed for 1 min with chemiluminescent detection reagent, according to the manufacturer's instructions. Exposure time was 10min. MW standards used, Page ruler, pre-stained from Thermofisher.
Courtesy of Dr. Göran Samuelsson, Umeå Plant Science Centre, Sweden - Additional Information
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Additional information: This antibody recognizes native and recombinant carbonic anhydrase of Chlamydomonas reinhardtii. - Background
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Background: CAH3 - carbonic anhydrase is a zinc-containing metalloenzyme that catalyzes the reversible interconversion of CO2 and HCO3?. It plays an important role in many physiological functions that involve decarboxylation or carboxylation reactions, including both photosynthesis and respiration.
- Product Citations
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Selected references: Terentyev and Shukshina (2024). CAH3 from Chlamydomonas reinhardtii: Unique Carbonic Anhydrase of the Thylakoid Lumen. Cells 2024, 13(2), 109.
Burlacot et al. (2022) Alternative photosynthesis pathways drive the algal CO2-concentrating mechanism. Nature 605, 366–371 (2022). https://doi.org/10.1038/s41586-022-04662-9
Shukshina &Terentyev (2021) Involvement of Carbonic Anhydrase CAH3 in the Structural and Functional Stabilization of the Water-Oxidizing Complex of Photosystem II from Chlamydomonas reinhardtii. Biochemistry Moscow 86, 867–877 (2021). https://doi.org/10.1134/S0006297921070075
Terentyev (2020: The Main Structural and Functional Characteristics of Photosystem-II-Enriched Membranes Isolated From Wild Type and cia3 Mutant Chlamydomonas reinhardtii. Life (Basel). 2020 May 14;10(5):E63. doi: 10.3390/life10050063.
Terentyev et al. (2019). Carbonic anhydrase CAH3 supports the activity of photosystem II under increased pH. Biochim Biophys Acta Bioenerg. 2019 Jul 1;1860(7):582-590. doi: 10.1016/j.bbabio.2019.06.003.
Correa-Galvis et al. (2016). Photosystem II Subunit PsbS Is Involved in the Induction of LHCSR Protein-dependent Energy Dissipation in Chlamydomonas reinhardtii. J Biol Chem. 2016 Aug 12;291(33):17478-87. doi: 10.1074/jbc.M116.737312.
Mitchell et al. (2014). Dynamics of carbon concentrating mechanism induction and protein re-localisation during the dark to light transition in synchronised Chlamydomonas. Plant Physiol. 2014 Aug 8. pii: pp.114.246918.
Tirumani et al. (2014). Regulation of CCM genes in Chlamydomonas reinhardtii during conditions of light-dark cycles in synchronous cultures. Plant Mol Biol. 2014 Mar 4.
Shutova et al. (2008). The photosystem II-associated Cah3 in Chlamydomonas enhances the O2evolution rate by proton removal. EMBO J. 27:782-791. - Protocols
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Agrisera Western Blot protocol and video tutorials
Protocols to work with plant and algal protein extracts - Reviews:
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James V Moroney | 2010-12-16We have ordered your CAH3 antibody AS05 073 and have been 100% happy with the specificity and sensitivity of this antibody.
Accessories
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