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NdhS | NAD(P)H-quinone oxidoreductase subunit S (chloroplastic)

AS16 4066 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana, Hordeum vulgare

NdhS | NAD(P)H-quinone oxidoreductase subunit S (chloroplastic) in the group Antibodies Plant/Algal  / Photosynthesis  / Electron transfer at Agrisera AB (Antibodies for research) (AS16 4066)
NdhS | NAD(P)H-quinone oxidoreductase subunit S (chloroplastic)



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Product Information

Immunogen

KLH-conjugated synthetic peptide dervied from Arabidopsis thaliana NdhS sequence, UniProt: A0A178V0R5, TAIR: AT4G23890

Host Rabbit
Clonality Polyclonal
Purity Immunogen affinity purified serum in PBS pH 7.4.
Format Lyophilized
Quantity 50 ĩg
Reconstitution For reconstitution add 50 ĩl 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 Western blot (WB)
Recommended dilution

1:10 000 (WB)

Expected | apparent MW

27 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana, Hordeum vulgare
Predicted reactivity Trema orientale, Parasponia andersonii, Populus tomentosa, Prunus avium, Raphanus sativus, Ziziphus jujuba
Species of your interest not listed? Contact us
Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples Application example

Western blot using anti-NdhS antibodies

Arabidopsis thaliana leaves from wildtype and ndho mutant were frozen in liquid nitrogen. Leaf tissue was homogenized in ice-cold isolation buffer (330 mm Suc, 25 mm HEPES-KOH, pH 7.4, 10 mm MgCl2, and 10 mm NaF) and filtered through Miracloth. The filtrate was centrifuged at 6,000g for 5 min at 4°C. The thylakoid pellet was resuspended in 25 mm HEPES-KOH, pH 7.4, 10 mm MgCl2, and 10 mm NaF, centrifuged at 6,000g, for 5 min at 4°C, and finally suspended in the isolation buffer and stored at −80°C. The chlorophyll content of isolated thylakoids was determined according to Porra et al. (1989)

Samples were denatured with Laemmli buffer at 65°C for 5 min. and were separated on  12 % SDS-PAGE  and blotted 1h to PVDF (0.45um), using semi-dry  transfer. Blot was blocked with 5 % milk 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 10000 in TBS-T ON/4°C with agitation. The antibody solution was decanted and the blot washed once for 15 min and 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) diluted to 1:25 000 in  for 1h/RT with agitation. The blot was washed as above and incubated for 5min min with chemiluminescent detection reagents. Exposure time was 5-10 min. 

Courtesy Dr. Lauri Nikkanen, University of Turku, Finland

Additional information

Related products

Background

Background

NAD(P)H-quinone oxidoreductase subunit 2 is involved in the transfer of electrons from NAD(P)H:plastoquinone, via FMN and iron-sulfur centers, to quinones in the photosynthetic chain and possibly in a chloroplast respiratory chain. Alternative names: NAD(P)H dehydrogenase, subunit 2, NADH-plastoquinone oxidoreductase subunit 2.

Product citations

Selected references Nikkanen et al. (2018). Regulation of cyclic electron flow by chloroplast NADPH‐dependent thioredoxin system. Plant Direct https://doi.org/10.1002/pld3.93
immunogen:

KLH-conjugated synthetic peptide dervied from Arabidopsis thaliana NdhS sequence, UniProt: A0A178V0R5, TAIR: AT4G23890

Reconstitution: For reconstitution add 50 ĩl of sterile water
Host: Rabbit
Clonality: Polyclonal
Purity: Immunogen affinity purified serum in PBS pH 7.4.
Format: Lyophilized
Quantity: 50 ĩg
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: Western blot (WB)
recommended dilution:

1:10 000 (WB)

calculated | apparent molecular mass [kDa]:

27 kDa

Confirmed reactivity: Arabidopsis thaliana, Hordeum vulgare
predicted reactivity: Trema orientale, Parasponia andersonii, Populus tomentosa, Prunus avium, Raphanus sativus, Ziziphus jujuba
Species of your interest not listed? Contact us
not reactive in: No confirmed exceptions from predicted reactivity are currently known
Picture (footer): Application example

Western blot using anti-NdhS antibodies

Arabidopsis thaliana leaves from wildtype and ndho mutant were frozen in liquid nitrogen. Leaf tissue was homogenized in ice-cold isolation buffer (330 mm Suc, 25 mm HEPES-KOH, pH 7.4, 10 mm MgCl2, and 10 mm NaF) and filtered through Miracloth. The filtrate was centrifuged at 6,000g for 5 min at 4°C. The thylakoid pellet was resuspended in 25 mm HEPES-KOH, pH 7.4, 10 mm MgCl2, and 10 mm NaF, centrifuged at 6,000g, for 5 min at 4°C, and finally suspended in the isolation buffer and stored at −80°C. The chlorophyll content of isolated thylakoids was determined according to Porra et al. (1989)

Samples were denatured with Laemmli buffer at 65°C for 5 min. and were separated on  12 % SDS-PAGE  and blotted 1h to PVDF (0.45um), using semi-dry  transfer. Blot was blocked with 5 % milk 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 10000 in TBS-T ON/4°C with agitation. The antibody solution was decanted and the blot washed once for 15 min and 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) diluted to 1:25 000 in  for 1h/RT with agitation. The blot was washed as above and incubated for 5min min with chemiluminescent detection reagents. Exposure time was 5-10 min. 

Courtesy Dr. Lauri Nikkanen, University of Turku, Finland
background:

NAD(P)H-quinone oxidoreductase subunit 2 is involved in the transfer of electrons from NAD(P)H:plastoquinone, via FMN and iron-sulfur centers, to quinones in the photosynthetic chain and possibly in a chloroplast respiratory chain. Alternative names: NAD(P)H dehydrogenase, subunit 2, NADH-plastoquinone oxidoreductase subunit 2.

All references: Nikkanen et al. (2018). Regulation of cyclic electron flow by chloroplast NADPH‐dependent thioredoxin system. Plant Direct https://doi.org/10.1002/pld3.93

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