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Agrisera Super Deal

Primary/Secondary/ECL



Add only 20 € to your primary antibody purchase
and you will also receive:

- Secondary antibody
Goat anti-rabbit, HRP conjugated
(to be used >1: 25 000)


- Two chemiluminescent detection reagents
Agrisera ECL Bright/SuperBright,  for 10 midi blots (picogram and femtogram detection range)

AS18 PrimarySecondaryECL

cAPX | Ascorbate peroxidase (cytosolic)

AS06 180  |  Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: A.  thaliana (immunohistochemistry only), A. toxicaria, D. salina, H.vulgare, P.sativum, S. lycopersicum, S. tuberosum, S. vulgaris, P. silvestris, . Z. mays

cAPX | Ascorbate peroxidase (cytosolic) in the group Plant/Algal Antibodies / Environmental Stress / Oxidative stress at Agrisera AB (Antibodies for research) (AS06 180)

PRODUCT INFORMATION IN PDF

Qty: 
355 €

Info: More information Read reviews
product information
Background

Ascorbate peroxidase (APX) is the enzyme catalyzing the ascorbate-dependent reduction of hydrogen peroxide. Ascorbate (AA) plays a key role in defense against oxidative stress and is particularly abundant in fruits and photosynthetic tissues. AA is found in every compartment of the plant cell including the apoplast.

Immunogen

KLH-conjugated peptide derived from N-terminal of Zea mays cytosolic APX Q41772

Host Rabbit
Clonality Polyclonal
Clone
Purity Total IgG
Format Lyophilized in PBS pH 7.4
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 tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.
Tested applications Immunoprecipitation (IP), Immunolocalization (IL), Western blot (WB)
Related products

AS06 184 | anti-AO | apoplastic ascorbate oxidase

AS09 384 | anti-AO | ascorbate oxidase

AS09 383 | anti-AO | ascorbate oxidase, biotinylated antibody

AS08 368 | anti-APX | ascorbate peroxidase

Plant protein extraction buffer

Secondary antibodies

Additional information Total IgG concentration is 3 µg/µl.
application information
Recommended dilution 2 µg (IP), 1 : 500 (IL), 1: 2000 - 1 : 10 000 (WB)
Expected | apparent MW

28 kDa

Confirmed reactivity Arabidopsis thaliana, Dunaliella salina, A. toxicaria, Hordeum vulgare, Pisum sativum, Silene vulgaris, Solanum lycopersicum, Solanum lycopersicum, Solanum tuberosum, Picea silvestris, Zea mays, Zygophyllum fabago
Predicted reactivity Brassica. juncea, Citrus sinensis, Fragaria ananassa, Gossypium hirsutum, Nicotiana tabacum, Solanum tuberosum, Sorghum bicolor, Pinus pinaster, Vitis vinifera
Not reactive in

Glycine max

Additional information
Selected references Ferrer et al. (2018). Differential Pb tolerance in metallicolous and non-metallicolous Zygophyllum fabago populations involves the strengthening of the antioxidative pathways. Environ & Exp Botany, Vo. 150, June 2018, Pages 141-151.
Aroca et al. (2015). S-sulfhydration: a new post-translational modification in plant systems. Plant Physiology March 2015 pp.00009.2015.
Terrile et al. (2014). Nitric oxide-mediated cell death is triggered by chitosan in F. eumartii spores. Pest Manag Sci. 2014 Apr 25. doi: 10.1002/ps.3814.
Tsaniklidis et al. (2013). L-Ascorbic acid metabolism in parthenocarpic and seeded cherry tomatoes. Plant Growth Regul,DOI 10.1007/s10725-013-9845-0. (Solanum lycopersicum, immunolocalization)

Application example

Western blot using anti-cAPX antibodies on tomato samples


40 µg of total protein from Solanum lycopersicum extracted with Extraction buffer (1x Hepes buffer, cOmplete Mini protease inhibitor cocktail (1 tablet/10ml), 5mM DTT) and denatured with 1x SDS-loading buffer at 95°C for 5 min. Samples were separated on 12 % SDS-PAGE and blotted 1h to PVDF using dry transfer. Blots were blocked with 5% milk in TBST for 45 min with agitation. Blot was then washed once with TBST, and then incubated in the primary antibody a-cAPX at a dilution of 1:2000 ON/4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602) diluted to 1:5000 in for 30min/RT with agitation. The blot was washed as above and developed for 2 min with ECL Prime (Solutions A and B Amersham ECL Prime Peroxide Solution, mixed in a 1:1 ratio). Exposure time was set to automatic.

Courtesy Dr. Nuria Sanchez Coll, CRAG, Spain

Western blot using anti-cAPX antibodies on A.thaliana samples

20 µg of total protein from Arabidopsis thaliana total (1), soluble (2) and membrane (3) fractions were prepared according to Planas-Marquès et al. (2016)     and denatured with 1x SDS-loading buffer at 95°C for 5 min. Samples were separated on 12 % SDS-PAGE and blotted 1h to PVDF using dry transfer. Blots were blocked with 12 % milk in TBST for 45 min with agitation. Blot was then washed once with TBST, and then incubated in the primary antibody a-cAPX at a dilution of 1:10 000 ON/4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in Agrisera matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, AS09 602) diluted to 1:5000 in for 30min/RT with agitation. The blot was washed as above and developed for 2 min with ECL Prime (Solutions A and B Amersham ECL Prime Peroxide Solution, mixed in a 1:1 ratio). Exposure time was set to automatic.

Courtesy Dr. Nuria Sanchez Coll, CRAG, Spain


 

(A) , (B)- control antibody, anti-PIN1, (C,D) immunolocalization using anti-cAPX antibodies. 

Courtesy of Dr. Taras Pasternak


 

immunolocalization using anti-cAPX antibodies
 
immunolocalization in high expressing neurons

Dissociated rat hippocampal neurons 14 DIV Transfection of FLAG- and Nuclear Export Signal-tagged APEX2 (variant of cAPX; FLAG-APEX2-NES). Fixation: 4% formaldehyde for 15 min RT, 0.1% TritonX-100 permeabilization 20 min RT, 5% FCS blocking 1 hr RT. All in PBS. Primary antibody: 1:200 rabbit anti-cAPX, 4°C incubated for 24h, 1:1000 mouse anti-FLAG, RT 1h. 
Secondary antibodies: 1:1000 anti-rabbit Alexa488, RT 1 h, 1:1000 anti-mouse Alexa561, RT 1 h.

Courtesy of Dr. Tony Cijsouw, Biederer lab, Department of Neuroscience Tufts University School of Medicine, USA

||| For other applications, usage on species other than stated above or any other questions, please use the LiveChat option or contact us at support@agrisera.com