Agrisera blog

Sample: Maize protoplasts fixed in 4% paraformaldehyde in 1X PBS pH 7.4 for 1 hour and washed 3 times in 1XPBS buffer

Primary antibodies: Agrisera AS21 4615 actin monoclonal; clone 14H4G8;

Secondary antibodies: Agrisera Donkey anti-Mouse IgG (H&L), DyLight® 488 conjugated AS10 1201

Other reagents: 30% Bovine Serum Albumin solution, Sigma Aldrich Cat # A8577-50ML; Phosphate Buffered Saline 10X, Electron Microscopy Sciences Cat # 19342-10; Imaging spacer, Millipore Sigma Grace Bio-Labs SecureSeal™ imaging spacer Cat # GBL654008

Protocol:

1. Block isolated protoplasts in a blocking buffer (2% BSA in 1x PBS) for 1 h at room temperature.
2. Dilute the primary antibody to 1: 500 in a blocking buffer, mix well, and spin down at 150 x g for 3 min:
3. Spin down protoplasts in a blocking buffer at 150 g for 1 -3 min and remove as much supernatant as possible using a pipette tip with a cut end (wide opening).
4. Resuspend protoplasts in diluted antibody and incubate for 1 h at room temperature.
5. Spin down protoplasts at 150 g for 1 min and remove as much supernatant as possible using a pipette tip with a cut end.
6. Wash protoplasts in a blocking buffer 3 times for 5 minutes each. Spin and resuspend as described in step 5 between each washing step.
7. Dilute the secondary antibody AS10 1201 to 1: 400, mix well, and spin down at 150 x g for 3 min:
8. Incubate protoplasts in diluted antibody for 1 h at room temperature.
9. Repeat the washing steps as described in 5-6.
10. Resuspend a protoplast pellet in a small volume of PBS 1X, pH 7.4.
11. Prepare microscopy slide: attach imaging spacer on top of the slide, peal off top adhesive membrane, add protoplast suspension on to the slide inside the spacer opening, cover with 22x22 mm cover glass No 1.5.
12. Imaging: Zeiss Elyra 7 SIM

Courtesy of Dr. Anastasiya Klebanovych, Dr. Kirk Czymmek, Dr. Kevin Cox, Dr. Blake Meyers, at the Donald Danforth Plant Science Center, USA

Agrisera supported the 5th International Plant Proteomics Organization (INPPO) conference, held between the 22nd and 25th of May 2023, in Thessaloniki, Greece. The conference focused on basic and applied plant proteomics research. 

The meeting covered the following topics:

• Plant stress under climate crisis
• Developmental proteomics
• Signaling in plant interactions
• Systems biology studies
• Technical/methodological advances in proteomic analysis

The winners of Agrisera Best Poster Prizes, as well as a free antibody of their choice from the Agrisera catalog, were Dr. Jingjing Huang from Department of Plant Biotechnology and Bioinformatics, Ghent University and VIB Center for Plant Systems Biology,Belgium. and PhD student Siel Goethals from VIB-UGent Plant Systems Biology, Belgium. 

Awaiting your free antibody choices!

• Actin (AS22 4615)
• Clathrin heavy chain 1,2 (AS10 690)
• Histone 3 (AS10 710A), and
• GFP (AS21 4443)

• Donkey anti-mouse IgG (H&L), DyLight® 488-conjugated secondary antibodies (AS10 1261), and
• Donkey anti-Rabbit IgG (H&L), DyLight® 550-conjugated secondary antibody (AS12 1962)

As climate change becomes more noticeable, and affect all life on Earth, research on how plants respond to environmental stress intensifies in many laboratories worldwide. Agrisera antibodies, to key proteins involved in plant stress response, help solve many pressing questions of how plants tackle changes to, and threats in, their environment. New antibodies are added continuously to our collection, which includes antibodies to proteins involved in:

Cold stress | Drought stress | Heat shock | Heavy metals | Hypoxia | Oxidative stress | Pathogen attack | Salt stress | UV radiation | Wounding

Research area

Agrisera supported the 32nd Western Photosynthesis Conference, held between the 28th and 30th of April 2023, in Bodega Biological Marine Laboratory, California, USA.

The meeting covered the following topics:

• Structure-Function Relations in Photosynthesis
• Electron Transfer and Organization of Photosynthetic Complexes
• Applied/Artificial Photosynthesis and Bioenergy
• Global Ecology and Photosynthetic Response to the Dynamic Environment
• Genomes and Evolution of Photosynthesis
• Carbon Fixation and Central Carbon Metabolism

The winners of Agrisera Best Talk Prizes, and a free antibody of their choice from the Agrisera catalog, were: 

For your convenience, chosen Agrisera antibodies can be purchased as sets at very favorabe pricing. The sets also include high-titer matching secondary antibodies, which can be used at 1: 25 000 1h/RT incubation, and Agrisera's ECL reagents, which offer versatile detection.

Agrisera offers the following antibody sets: 

Educational antibody set for photosynthesis | Photosynthesis Tool kit – quantitation (includes standards for quantification of PSI and PSII) | Oxidative stress antibody set | Argonaute antibody set | Rubisco quantitation set (for Western blot or ELISA) | Plant Cell Compartment Antibody Marker Set for 3, 5 or 10 marker antibodies of your choice.

You are always welcome to suggest a new antibody set to us!

Antibodies to Arabidopsis thaliana proteins involved in photosynthesis are a core collection of the Agrisera catalog. This group of products has been developed in collaboration with research laboratories worldwide, for the last 20 years. Agrisera photosynthetic antibodies have the following features:

• Developed to well-conserved peptides, which allows for broad species reactivity
• Can be used at high dilutions, which allows many tests to be conducted
• Validated in independent laboratories, with appropriate sets of endogenous samples (no recombinant protein validation only)
• Available since the year 2000, meaning this collection is prided by an extensive publication record
• For your convenience, some of the antibodies to major photosynthetic proteins, are put together into sets, at discounted pricing. In some cases, these are also accompanied by quantitation standards, to allow estimation of PSI/PSII ratio.

Beside antibodies, Agrisera has also developed Educational Posters on Photosynthesis, that are engaging and packed with up-to-date information. These posters are available for download, and as hard copies that can be requested here. Each poster was created in consultation with leading researchers from the scientific community, and the information in the posters is updated regularly. Specific figures and schemes from the posters are available free of charge for download, to be used in publications and presentations.

Samples: 

1: 13.5 µg of Eucalyptus grandis whole leaf extract clone one (g1)
4: 13.5 µg of Eucalyptus grandis whole leaf extract clone two (g2)
6: 13.5 µg of one clone Eucalyptus grandis x Eucalyptus camaldulensis whole leaf extract (gc)
8: 13.5 µg of one clone Eucalyptus grandis x Eucalyptus tereticornis whole leaf extract (gt)

Presented data courtesy of Ing. Agr. MSc. Matías Nión from Departamento de Biología Vegetal, Facultad de Agronomía, UdelaR. Montevideo, Uruguay.

With millions of antibodies offered worldwide, by hundreds of suppliers, it may be a bit overwhelming, especially for beginners, to choose the correct antibody to answer a specific questions in a research project. Here is a short checklist with what to pay attention to:

Species reactivity – Has the antibody been used to detect the protein from the species we are working with? If not, a request should be sent to the supplier to confirm if the antibody may have so-called predicted reactivity to the target protein. If not, the antibody is not suitable, and false positive results can be produced.  

Validation data – Is validation data available on a product information sheet? For non-essential proteins, validation data should include results obtained with knockdown or knockout mutants. Western blot results showing that an antibody binds to a recombinant protein, are not relevant to the binding in a specific sample, extracted from a specific tissue or organism. Here we explain why.

Application – If the antibody has only been used in Western blot, it may not work in immunoprecipitation or immunolocalization. Each of these techniques requires separate validation data.

Recommended antibody dilution – The lower the dilution, the fewer tests can be conducted using an antibody. Here we explain why. 

Publication record – Being citet in scientific articles definitely confirms the performance of a given antibody. However, as there is no worldwide standard, product numbers are oftentimes not included in the "Material and Method" section. This makes it difficult to identify what antibody has been used to obtain the presented results. How to cite antibodies in scientific publication, is explained in this short video.

Well-validated antibodies to specific cellular compartments are crucial to support data regarding localization studies of novel proteins. Agrisera's Cellular compartment antibody collection contains of marker antibodies to:

Cell wall | Chloroplast | Cytoplasm | Microsomal membranes | Mitochondria | Nuclei | Peroxisomes | Plasma membrane | Vacuole

Agrisera compartment marker antibodies have been used in in a wide range of publications over the last 15 years. For many compartments we offer several antibodies, to proteins of different molecular weight, to match your experimental requirements.

Not certain which compartment marker antibody to choose? Ask Agrisera's techincal staff for an advice.  


Until end of June 2023, Agrisera Antibody Collection for Arabidopsis thaliana is provided with a 10% discount. 

Agrisera offers an extensive and well-published collection of antibodies to support research on the biology of the model plant Arabidopsis thaliana.

Advantages of this collection include:

• Validation data, often provided with Agrisera high-titer matching secondary antibodies and outstanding detection reagents
• Antibodies detecting proteins involved in various processes, and specific cellular compartments (compartment markers)
• Extensive publication record. Some of Agrisera antibodies have been available for over 20 years.
• Minimal lot-to-lot variation
• Provided in a safe and economical format (lyophilized) for environmentally-friendly shipments that do not require heavy packages or dry ice
• Prompt technical support

Find a matching antibody to a gene ID | Complete list of antibodies with confirmed and predicted reactivity to Arabidopsis thaliana | Examples of publications with Agrisera Antibodies | Benefit from a 10% discount until end of June with the discount code Arabi10

Extensive collection of antibodies to Arabidopsis thaliana proteins involved in photosynthesis

Extensive collection of antibodies to Arabidopsis thaliana proteins involved in a stress response

Agrisera's Arabidopsis thaliana antibodies are suitable for a wide range of techniques, including immunofluorescence, ChIP, IP and Western Blot. Over 100 antibodies are available free of charge for testing, in return for results. Check the list here. 

• High sensitivity
• High average dilution antibody can be used at
• Long-term stability
  
• Superior lot-to-lot consistency
• Most of Agrisera primary antibodies were validated in pair with Agrisera secondary antibodies 

 

Poster price award, 1st place. From left to right: Trang Vuong, Prof. Dr. Andreas Holzinger. Photograph by: PD Dr. Volker Wagner

• High sensitivity
• Long-term stability
  
• Suitable for PVDF and nitrocellulose membranes
• Superior lot-to-lot consistency
• Competitive pricing

Glutamine synthetase GlnA is the key enzyme in the incorporation of mineral nitrogen into glutamine. The activity of this enzyme is controlled by adenylarion under conditions of abundant glutamine.

Agrisera Global GlnA antibodies are reactive to this enzyme from the following species, as confirmed experimentally: Deinococcus radioduransm Synechococcus sp. strain PCC 7942, Synechocystis sp. strain PCC 6803, Trichodesmium IMS

Confirmed and predicted reactivity is explained further here. 

By using GlnA Global antibodies, combined with GlnA protein standard it is possible to quantify GlnA using quantitative Western blot, described here, and in this video tutorial.

• The antigen used to elicit the antibody
• Confirmed and predicted reactivity of the antibody
• Applications and recommended starting dilution
• Recently published articles

• Plant cell wall properties and dynamics
• Epigenetic regulation
• Population genetics
• Omics approaches
• Plant nutrition
• Development and Signaling
• Response to mechanical stress
• Adaptation to abiotic stress 

Nitrogenase NifH is involved in the biological fixation of atmospheric nitrogen to ammonia. Alternative protein names are Nitrogenase component II, Nitrogenase Fe protein, Nitrogenase reductase, and FeMoCo-nitrogenase.

Agrisera Global anti-NifH antibodies are reactivie to this enzyme from the following species, as confirmed experimentally: Anabaena PCC7120, Clostridium butyricum, Codakia orbicularis, Cylindrospermopsis raciborskii CS-505, Dolichospermum sp., Nostoc sp, Rhodopseudomonas palustris, Trichodesmium sp., nodules of Trifolium repens L., Vibrio natriegens ATCC 14048

Confirmed and predicted reactivity is explained further here. 

By using NifH antibodies, combined with NifH protein standard it is possible to quantify the enzyme using quantitative Western blot, described here, and in this video tutorial.

Agrisera goat anti-rabbit IgG secondary antibodies, conjugated to horseradish peroxidase (HRP), perform in dilution > 1: 25 000 in Western blot and ELISA, which is supported by a long list of publications.

As this anti-rabbit IgG secondary antibody performs in high dilutions, it offers advantages to other secondary antibodies in terms of minimizing background signal, and the number of experiments it can be used for. This makes it very economical, and strengthens experimental reproducibility.

Additionally, this antibody is provided in lyophilized format, which makes shipment economical and environmentally friendly, as there is no need to include dry ice. Shipments are done at ambient temperature, with perserved antibody performance, which has been tested and evaluated over the last 20 years.

Until the end of 2023, this product is offered at a 10% discount using the discount code: Conj10.

Secondary antibodies conjugated to HRP (Horseradish peroxidase) and ALP (Alkaline phosphatase) enzymes are most frequently used to visualize target proteins in Western blot, ELISA and immunolocalization. When using secondary antibodies conjugated to HRP, the detection is performed using chemiluminescence (recorded by a CCD camera or another imager), which gives high sensitivity and good resolution in both Western blot and chemiluminescent ELISA. When using antibodies conjugated to ALP, visualization of the antibody binding to the target is achieved using chromogenic reagents, and no CCD camera is necessary.  

Agrisera's secondary antibody collection is composed of over 220 antibodies; both highly purified and available in a basic format, suitable for Western blot, as well as adsorbed to serum from a wide range of animal species, suitable for immunolocalization and ELISA. Based on the analysis of antibody performance in available publications, conducted by CiteAB, it became very clear that Agrisera secondary antibodies are performing above market average. This means that Agrisera secondary antibodies conjugated to ALP or HRP can be used at high dilutions and are price-worthy reagents, and will provide you with a consistent performance over many experiments.

Broad species reactivity

• Broad species reactivity, including both di- and monocols and microalgae
• Can be used in dilutions of >1: 1000 in Western blot
• Extensive publication record
• Promptly and environmentally-friendly delivery

Agrisera will participate in the following meetings in 2023:

3^rd INUPRAG Symposium on Integrative Plant Biology, 21^st-22^nd of February, Umeå Plant Science Centre, Umeå, Sweden
The 33^rd International Conference on Arabidopsis Research, 5^th-9^th of June, Chiba, Japan
Nordic Photosynthesis Congress and Nordic Algae Symposium, 19^th-21^st of June, Umeå, Sweden
Plant Biology Europe, 3^rd-6^th of July, Marseille, France
Plant Biology 2023 (ASPB), 5^th-8^th of August, Savannah, Georgia, USA
11^th Conference of Polish Society of Experimental Plant Biology, 19^th-22^nd of September, Poznań, Poland
KBC Days 2023, November, Umeå University, Sweden 

Agrisera supports the following meetings in 2023:

Umeå Center for Microbial Research Day (UCMR Day 2023), 19^th of January, Umeå University, Sweden
36^th Molecular Biology of Plants (MBP2023), 6^th-9^th of February, Bonn, Germany
20^th Scientific Conference of the Phycology Section, 12^th-14^th of March, Jena, Germany
19th International Workshop on Plant Membrane Biology, 27^th-31^st of March, Taipei, Taiwan
ATSPB 2023: Plants in the context of food security, biodiversity and climate change, 13^th-15^th of April, Tirol, Austria
32^nd Western Photosynthesis Conference, 28^th-30^th of April, Bodega Bay, California, USA
5^th Conference of the International Plant Proteomics Organization, 22^nd-25^th of May, Thesaloniki, Greece
IPG 2023 Symposium - Redox Regulation of Plant Stress and Development, 23^th - 26^th of May, University of Missouri, USA
6^th UNIGE PostDoc Day, 8^th of June at the University of Geneva, Switzerland
Chlamy2023, The 20th International Conference on the Cell and Molecular Biology of Chlamydomonas, 4^th-9^th of June, Princeton University, New Jersey, USA
Plant Biology 2023, XXV Meeting of the Spanish Society of Plant Biology, 9^th-12^th of July, Braga, Portugal
4^th International Brassinosteroid Conference, 16^th-18^th of August, Ghent, Belgium
Plants and People, 6t^h-7^th of September, Max Planck Institute of Molecular Plant Physiology, Potsdam, Germany
Research Day, 20^th of September, Oslo, Norway
11^th Conference of Polish Society of Experimental Plant Biology, 19^th-22^nd of September, Poznań, Poland
2^nd International Conference on Plant Systems Biology and Biotechnology (ICPSBB), 25^th-30^th of September, Plovdiv, Bulgaria
12^th SPPS PhD conference PhD 101: Everything is hard before it gets easy, 27^th-29^th of September, Lund, Sweden
4^th Annual Congress of Young Researchers (ACYR), 9^th-10^th of October, CRAG auditorium, Spain
49th Midwest/Southeast Photosynthesis Conference, , 27^th-29^th of October, Turkey Run State Park, USA
The Canadian Society of Plant Biologists (CSPB), Eastern Regional Meeting, 1^st-2^nd of December, Concordia University, Montreal, Canada.

 

In the image, left to right: Dr. Rikesh Bhalerao (UPSC), Dr. Sara Raggi (UPSC) and Dr. Joanna Porankiewicz-Asplund (Agrisera).
 
"The prize gives us the opportunity to thank our personnel for their work and commitment. It is always very difficult to choose between the candidates, as they all highly deserve this prize," says Rishikesh Bhalerao who announced the winner of the prize this year.

Photo by Prof. Stephanie Robert.