ASN | Glutamine-dependent asparagine synthetase

AS20 4425 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

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ASN | Glutamine-dependent asparagine synthetase

Product Information

Immunogen Purified full length, tag cleaved, recombinant Arabidopsis thaliana ASN2, UniProt: Q9LV77 , TAIR: AT5G65010

Host Rabbit

Clonality Polyclonal

Purity Total IgG. Protein A purified in PBS, 50% glycerol. Filter sterilized.

Format Liquid at 2 mg/ml.

Quantity 200 µg

Storage Store 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 ELISA (ELISA), Immunohistochemistry (IHC), Western blot (WB)

Recommended dilution assay dependent (ELISA), 1: 100-1: 500, paraffin sections (IHC), 1: 1000-1: 2000 (WB)

Expected | apparent MW 65 | 65 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana, Zea mays

Predicted reactivity Brassica rapa, Camelina sativa, Capsella rubella, Eutrema salsugineum, Punica granatum
Species of your interest not listed? Contact us

Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples Western blot using anti-ASN Glutamine-dependent asparagine synthetase

Arabidopsis thaliana total leaf extract was freshly extracted with 2x SDS-sample buffer (+ 2ME) for SDS-PAGE and denatured with 4X SDS buffer at 95°C for 5 min. Protein was loaded/well and were separated on 10% SDS-PAGE and blotted 1h to PVDF membrane. Blot was blocked with 3 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 in TBS-T for 1h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendation. Molecular weight of ASN2 is 65 kDa. ASN1 is expressed in floral organs, while ASN2 is expressed in leaf.

Western blot using anti-ASN antibodies on wilde type and mutants

Western blot using anti-ASN antibodies on wilde type and mutants

Arabidopsis thaliana total leaf extract and respective mutants were freshly extracted with 2x SDS-sample buffer (+ 2ME) for SDS-PAGE and denatured with 4X SDS buffer at 95°C for 5 min. 10 µg of protein was loaded/well abd samples were separated on 10% SDS-PAGE and blotted 1h to PVDF membrane. Blot was blocked with 3 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2500 in TBS-T for 1h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendation.

For images of sections and IHC method protocol, please refer for listed publications.

Additional information

Additional information This antibody reacts with both isoforms: ASN1 and ASN2

Background

Background ASN (Glutamine-dependent asparagine synthetase) is essential for nitrogen assimilation, distribution and remobilization within the plant via the phloem. ASN2 is expressed in leaf and ASN1 is expressed in floral organs. The amino acid sequences of Arabidopsis thaliana ASN1 and ASN2 are 76% identical. The amino acid sequences of Arabidopsis thaliana and Zea mays ASN2 are 73.6% identical.

Product citations

Selected references Gaufichon et al. (2017). ASN1-encoded asparagine synthetase in floral organs contributes to nitrogen filling in Arabidopsis seeds. Plant J. 2017 Aug;91(3):371-393. doi: 10.1111/tpj.13567.
Gaufichon et al. (2013). Arabidopsis thaliana ASN2 encoding asparagine synthetase is involved in the control of nitrogen assimilation and export during vegetative growth. Plant Cell Environ. 2013 Feb;36(2):328-42. doi: 10.1111/j.1365-3040.2012.02576.x.

background:	ASN (Glutamine-dependent asparagine synthetase) is essential for nitrogen assimilation, distribution and remobilization within the plant via the phloem. ASN2 is expressed in leaf and ASN1 is expressed in floral organs. The amino acid sequences of Arabidopsis thaliana ASN1 and ASN2 are 76% identical. The amino acid sequences of Arabidopsis thaliana and Zea mays ASN2 are 73.6% identical.
All references:	Gaufichon et al. (2017). ASN1-encoded asparagine synthetase in floral organs contributes to nitrogen filling in Arabidopsis seeds. Plant J. 2017 Aug;91(3):371-393. doi: 10.1111/tpj.13567. Gaufichon et al. (2013). Arabidopsis thaliana ASN2 encoding asparagine synthetase is involved in the control of nitrogen assimilation and export during vegetative growth. Plant Cell Environ. 2013 Feb;36(2):328-42. doi: 10.1111/j.1365-3040.2012.02576.x.
additional information:	This antibody reacts with both isoforms: ASN1 and ASN2
calculated \| apparent molecular mass [kDa]:	65 \| 65 kDa
Clonality:	Polyclonal
Format:	Liquid at 2 mg/ml.
Host:	Rabbit
immunogen:	Purified full length, tag cleaved, recombinant Arabidopsis thaliana ASN2, UniProt: Q9LV77 , TAIR: AT5G65010
Purity:	Total IgG. Protein A purified in PBS, 50% glycerol. Filter sterilized.
Quantity:	200 µg
recommended dilution:	assay dependent (ELISA), 1: 100-1: 500, paraffin sections (IHC), 1: 1000-1: 2000 (WB)
storage:	Store 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:	ELISA (ELISA), Immunohistochemistry (IHC), Western blot (WB)
Confirmed reactivity:	Arabidopsis thaliana, Zea mays
predicted reactivity:	Brassica rapa, Camelina sativa, Capsella rubella, Eutrema salsugineum, Punica granatum Species of your interest not listed? Contact us
not reactive in:	No confirmed exceptions from predicted reactivity are currently known
Picture (footer):	Arabidopsis thaliana total leaf extract was freshly extracted with 2x SDS-sample buffer (+ 2ME) for SDS-PAGE and denatured with 4X SDS buffer at 95°C for 5 min. Protein was loaded/well and were separated on 10% SDS-PAGE and blotted 1h to PVDF membrane. Blot was blocked with 3 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1000 in TBS-T for 1h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendation. Molecular weight of ASN2 is 65 kDa. ASN1 is expressed in floral organs, while ASN2 is expressed in leaf. Arabidopsis thaliana total leaf extract and respective mutants were freshly extracted with 2x SDS-sample buffer (+ 2ME) for SDS-PAGE and denatured with 4X SDS buffer at 95°C for 5 min. 10 µg of protein was loaded/well abd samples were separated on 10% SDS-PAGE and blotted 1h to PVDF membrane. Blot was blocked with 3 % skim milk/TBS-T, 1h/RT with agitation. Blot was incubated in the primary antibody at a dilution of 1: 2500 in TBS-T for 1h/RT. The antibody solution was decanted and the blot was washed 4 times for 10 min in TBS-T at RT with agitation. Blot was incubated in matching secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated) diluted to 1:10 000 in for 1h/RT with agitation. The blot was washed as above and developed with a chemiluminescent detection reagent, following manufacture's recommendation. For images of sections and IHC method protocol, please refer for listed publications.