PHOT1 | Phototropin-1

AS10 720 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from known Arabidopsis thaliana PHOT1 O48963, At3g45780

Host Rabbit

Clonality Polyclonal

Purity Serum

Format Lyophilized

Quantity 50 µl

Reconstitution For reconstitution add 50 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 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 : 5000 (WB)

Expected | apparent MW

111 | 132 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana

Predicted reactivity Arabidopsis thaliana

Not reactive in Cuscuta campestris, Oryza sativa

Application examples

Application examples application example
western blot using anti-PHOT1 antibodies

80 µg of total protein from Arabidopsis thaliana wt Columbia (right lane) and Arabidopsis thaliana phot1phot2 double mutant (left lane) were separated on 9% SDS-PAGE and blotted 2h to PVDF. Blots were blocked immediately following transfer in PBS-T 5% milk for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 5000 overnight at 4^oC with agitation. The antibody solution was decanted and the blot was rinsed briefly, then washed 3 times for 5 min in PBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:10 000 for 1h at room temperature with agitation. The blots were washed as above and developed with WestPico detection reagent (PIERCE) according to the manufacturers instructions. Exposure time was 300 seconds.

Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 27406783

Journal: J Exp Bot

Figure Number: 6C

Published Date: 2016-09-01

First Author: Sztatelman, O., ?abuz, J., et al.

Impact Factor: 6.088

Open Publication

Profiles of phototropin1 (A) and phototropin2 (B) expression in darkened and light-exposed (120 µmol m?2 s?1 and 3h) Arabidopsis wild-type and mutant (phot1, phot2, and rcn1) leaves at the mRNA level. Each point represents the average obtained from at least nine leaves of different plants. Error bars show the SE. Asterisks indicate statistically significant differences between samples *P=0.01–0.05. (C and D) A representative western blot showing the expression of PHOT1 (C) and PHOT2 (D) in wild-type and mutant plants. Proteins stained with CBB are shown as the well loading reference.

Reactant: Arabidopsis thaliana (Thale cress)

Application: Western Blotting

Pudmed ID: 27406783

Journal: J Exp Bot

Figure Number: 7A

Published Date: 2016-09-01

First Author: Sztatelman, O., ?abuz, J., et al.

Impact Factor: 6.088

Open Publication

Representative dephosphorylation profiles of phototropin1 after blue light exposure (120 µmol m?2 s?1 and 1h) in Arabidopsis wild-type and mutant (phot2 and rcn1) leaves. Dark, a dark-adapted control; 0, a sample collected just after illumination. The duration of the incubation in the darkness after the end of the illumination is indicated in minutes. Phosphorylation leads to the shift of the phototropin band towards higher mass. Samples treated with alkaline phosphatase are shown on the right. Anti-actin blots are presented as the loading reference. The results represent two out of 4–5 independent biological replicates.

Additional information

Additional information This product can be sold containing ProClin if requested

Information about phot1 mutant, first named nph1:

Liscum &. Briggs (1995). Mutations in the NPH1 Locus of Arabidopsis Disrupt the Perception of Phototropic Stimuli. The Plant Cell, Vol. 7, 473-485.
Huala et al. (1997). Arabidopsis NPH1: A Protein Kinase with a Putative Redox-Sensing Domain. Science 19: Vol. 278 no. 5346 pp. 2120-2123.
Lehmann et al. (2011). Transitions of gene expression induced by short-term blue light. Plant Biology Volume 13, Issue 2, pages 349–361. Seeds of this mutant are available at uNASC.

Background

PHOT1 | phototropin-1 is a blue-light photoreceptor which containst light activated serine-threonine kinase domain. Is required for stomatal opening, chloroplast movements, leaf flattening and phototropism. Undergoes blue-light-dependent autophosphorylation. Alternative names: NPH1, RPT1, JK224, Non-phototropic hypocotyl protein 1, Root phototropism protein 1

Product citations

Selected references Labuz et al. (2021) Phototropin interactions with SUMO proteins. Plant Cell Physiol. 2021 Feb 17:pcab027. doi: 10.1093/pcp/pcab027. Epub ahead of print. PMID: 33594440.
Krzeszowiec et al. (2020). Chloroplasts in C3 grasses move in response to blue-light. Plant Cell Rep . 2020 Oct;39(10):1331-1343.doi: 10.1007/s00299-020-02567-3. Epub 2020 Jul 13.
Labuz et al. (2015). The impact of temperature on blue light induced chloroplast movements in Arabidopsis thaliana. Plant Science, doi:10.1016/j.plantsci.2015.07.013.
Eckstein et al. (2015). Auxin and chloroplast movements. Physiol Plant. 2015 Oct 15. doi: 10.1111/ppl.12396.

immunogen:	KLH-conjugated synthetic peptide derived from known Arabidopsis thaliana PHOT1 O48963, At3g45780
Reconstitution:	For reconstitution add 50 µl of sterile water
Host:	Rabbit
Clonality:	Polyclonal
Purity:	Serum
Format:	Lyophilized
Quantity:	50 µl
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 : 5000 (WB)
calculated \| apparent molecular mass [kDa]:	111 \| 132 kDa
Confirmed reactivity:	Arabidopsis thaliana
predicted reactivity:	Arabidopsis thaliana
not reactive in:	Cuscuta campestris, Oryza sativa
Picture (footer):	application example 80 µg of total protein from Arabidopsis thaliana wt Columbia (right lane) and Arabidopsis thaliana phot1phot2 double mutant (left lane) were separated on 9% SDS-PAGE and blotted 2h to PVDF. Blots were blocked immediately following transfer in PBS-T 5% milk for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 5000 overnight at 4^oC with agitation. The antibody solution was decanted and the blot was rinsed briefly, then washed 3 times for 5 min in PBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:10 000 for 1h at room temperature with agitation. The blots were washed as above and developed with WestPico detection reagent (PIERCE) according to the manufacturers instructions. Exposure time was 300 seconds.
More images:	Reactant: Arabidopsis thaliana (Thale cress) Application: Western Blotting Pudmed ID: 27406783 Journal: J Exp Bot Figure Number: 6C Published Date: 2016-09-01 First Author: Sztatelman, O., ?abuz, J., et al. Impact Factor: 6.088 Open Publication Profiles of phototropin1 (A) and phototropin2 (B) expression in darkened and light-exposed (120 µmol m?2 s?1 and 3h) Arabidopsis wild-type and mutant (phot1, phot2, and rcn1) leaves at the mRNA level. Each point represents the average obtained from at least nine leaves of different plants. Error bars show the SE. Asterisks indicate statistically significant differences between samples P=0.01–0.05. (C and D) A representative western blot showing the expression of PHOT1 (C) and PHOT2 (D) in wild-type and mutant plants. Proteins stained with CBB are shown as the well loading reference. Reactant: Arabidopsis thaliana (Thale cress)* Application: Western Blotting Pudmed ID: 27406783 Journal: J Exp Bot Figure Number: 7A Published Date: 2016-09-01 First Author: Sztatelman, O., ?abuz, J., et al. Impact Factor: 6.088 Open Publication Representative dephosphorylation profiles of phototropin1 after blue light exposure (120 µmol m?2 s?1 and 1h) in Arabidopsis wild-type and mutant (phot2 and rcn1) leaves. Dark, a dark-adapted control; 0, a sample collected just after illumination. The duration of the incubation in the darkness after the end of the illumination is indicated in minutes. Phosphorylation leads to the shift of the phototropin band towards higher mass. Samples treated with alkaline phosphatase are shown on the right. Anti-actin blots are presented as the loading reference. The results represent two out of 4–5 independent biological replicates.
additional information:	This product can be sold containing ProClin if requested
additional information (application):	*Information about phot1* mutant, first named nph1:** Liscum &. Briggs (1995). Mutations in the NPH1 Locus of Arabidopsis Disrupt the Perception of Phototropic Stimuli. The Plant Cell, Vol. 7, 473-485. Huala et al. (1997). Arabidopsis NPH1: A Protein Kinase with a Putative Redox-Sensing Domain. Science 19: Vol. 278 no. 5346 pp. 2120-2123. Lehmann et al. (2011). Transitions of gene expression induced by short-term blue light. Plant Biology Volume 13, Issue 2, pages 349–361. Seeds of this mutant are available at uNASC.
background:	PHOT1 \| phototropin-1 is a blue-light photoreceptor which containst light activated serine-threonine kinase domain. Is required for stomatal opening, chloroplast movements, leaf flattening and phototropism. Undergoes blue-light-dependent autophosphorylation. Alternative names: NPH1, RPT1, JK224, Non-phototropic hypocotyl protein 1, Root phototropism protein 1
All references:	Labuz et al. (2021) Phototropin interactions with SUMO proteins. Plant Cell Physiol. 2021 Feb 17:pcab027. doi: 10.1093/pcp/pcab027. Epub ahead of print. PMID: 33594440. Krzeszowiec et al. (2020). Chloroplasts in C3 grasses move in response to blue-light. Plant Cell Rep . 2020 Oct;39(10):1331-1343.doi: 10.1007/s00299-020-02567-3. Epub 2020 Jul 13. Labuz et al. (2015). The impact of temperature on blue light induced chloroplast movements in Arabidopsis thaliana. Plant Science, doi:10.1016/j.plantsci.2015.07.013. Eckstein et al. (2015). Auxin and chloroplast movements. Physiol Plant. 2015 Oct 15. doi: 10.1111/ppl.12396.

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