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PAL 1-4 | Phenylalanine ammonia-lyase 1-4

AS21 4614   | Clonality:  Polyclonal |  Host:  Rabbit | Reactivity: Nicotiana benthamiana

PAL 1-4 | Phenylalanine ammonia-lyase 1-4 in the group Antibodies Plant/Algal  / Environmental Stress / Pathogen attack at Agrisera AB (Antibodies for research) (AS21 4614)
PAL 1-4 | Phenylalanine ammonia-lyase 1-4



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Product name, number (Agrisera, Sweden)

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Product Information

Immunogen KLH-conjugated, conserved peptide derived from Arabidopsis thaliana PAL1-4, UniProt:  P35510, P45724, P45725, Q9SS45 , TAIR:AT2G37040, AT3G53260, AT5G04230,AT3G10340
Host Rabbit
Clonality

Polyclonal

Purity Antigen affinity purified serum, in PBS pH 7.4
Format Lyophilized
Quantity 50 µg
Reconstitution For reconstitution add 50 µl, of sterile or deionized 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 Western blot (WB)
Recommended dilution 1 : 2000 (WB)
Expected | apparent MW 76.2-78.7 kDa

Reactivity

Confirmed reactivity Nicotiana benthamiana
Predicted reactivity Arabidopsis thaliana, Hibiscus syriacus, Lotus corniculatus, Solanum dulcamara, Solanum lycopersicum,Vigna unguiculata, 
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 phenylalanine ammonia-lyase 1-4

Target MW: ~77 kDa

Target cellular localisation: Cytoplasmic


~25 µg/well of total protein extracted freshly from N. benthamiana leaf. Sample 3 is a control, no overexpression. Exact buffer components were: Laemmli buffer (62.5mM Tris-HCl (pH 6.8), 10% glycerol, 1%SDS, 0.005% Bromophenol Blue): and denatured with exact buffer components at 70°C for 10 min.  Samples were separated on 10 % SDS-PAGE and blotted for 0.5 h to PVDF using semi-dry transfer. Blot was blocked with 5 % milk for 1h at RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:2 000 in TBS-T ON/4°C with agitation. The antibody solution was decanted, and the blot was rinsed briefly twice, then washed once for 15 min and 3 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 following chemiluminescent detection reagent: AS16 ECL-N-10 AgriseraBright (mid picogram detection). Exposure time was 5 minutes (LI-COR Odyssey FC Imaging System).

Image courtesy of Dr Yasin Tumtas, Bozkurt Lab, Imperial College London

Additional information

Related products

Background

Background This is a key enzyme of plant metabolism catalyzing the first reaction in the biosynthesis from L-phenylalanine of a wide variety of natural products based on the phenylpropane skeleton.

Product citations

immunogen: KLH-conjugated, conserved peptide derived from Arabidopsis thaliana PAL1-4, UniProt:  P35510, P45724, P45725, Q9SS45 , TAIR:AT2G37040, AT3G53260, AT5G04230,AT3G10340
Reconstitution: For reconstitution add 50 µl, of sterile or deionized water.
Host: Rabbit
Clonality:

Polyclonal

Purity: Antigen affinity purified serum, in PBS pH 7.4
Format: Lyophilized
Quantity: 50 µg
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: Western blot (WB)
recommended dilution: 1 : 2000 (WB)
calculated | apparent molecular mass [kDa]: 76.2-78.7 kDa
Confirmed reactivity: Nicotiana benthamiana
predicted reactivity: Arabidopsis thaliana, Hibiscus syriacus, Lotus corniculatus, Solanum dulcamara, Solanum lycopersicum,Vigna unguiculata, 
Species of your interest not listed? Contact us
not reactive in: No confirmed exceptions from predicted reactivity are currently known
Picture (footer): Western blot using anti phenylalanine ammonia-lyase 1-4

Target MW: ~77 kDa

Target cellular localisation: Cytoplasmic


~25 µg/well of total protein extracted freshly from N. benthamiana leaf. Sample 3 is a control, no overexpression. Exact buffer components were: Laemmli buffer (62.5mM Tris-HCl (pH 6.8), 10% glycerol, 1%SDS, 0.005% Bromophenol Blue): and denatured with exact buffer components at 70°C for 10 min.  Samples were separated on 10 % SDS-PAGE and blotted for 0.5 h to PVDF using semi-dry transfer. Blot was blocked with 5 % milk for 1h at RT with agitation. Blot was incubated in the primary antibody at a dilution of 1:2 000 in TBS-T ON/4°C with agitation. The antibody solution was decanted, and the blot was rinsed briefly twice, then washed once for 15 min and 3 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 following chemiluminescent detection reagent: AS16 ECL-N-10 AgriseraBright (mid picogram detection). Exposure time was 5 minutes (LI-COR Odyssey FC Imaging System).

Image courtesy of Dr Yasin Tumtas, Bozkurt Lab, Imperial College London
background: This is a key enzyme of plant metabolism catalyzing the first reaction in the biosynthesis from L-phenylalanine of a wide variety of natural products based on the phenylpropane skeleton.

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