GPA1 | Guanine nucleotide-binding protein subunit alpha 1

Name: GPA1 | Guanine nucleotide-binding protein subunit alpha 1
SKU: AS16 3939
Price: 302 EUR
Availability: InStock

AS16 3939 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana

GPA1 | Guanine nucleotide-binding protein subunit alpha 1 in the group Antibodies Plant/Algal / Plant Developmental Biology / Plant Signal Transduction at Agrisera AB (Antibodies for research) (AS16 3939)

GPA1 | Guanine nucleotide-binding protein subunit alpha 1

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from Arabidopsis thaliana GPA1, UniProt: P18064, TAIR: AT2G26300

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: 2000 (WB)

Expected | apparent MW 49,3 kDa

Reactivity

Confirmed reactivity Arabidopsis thaliana

Predicted reactivity

Amborella trichopoda, Ananas comosus, Arachis sp., Beta vulgaris, Brassica sp., Cajanus cajan, Camelina sativa, Capsella rubella, Capsicum annuum, Cicer arietinum, Coffea canephora, Cucumis melo, Daucus carota, Elaeis guineensis, Eucalyptus grandis, Eutrema sp., Fragaria vesca, Genlisea aurea, Glycine max, Glycine soja, Jatropha curcas, Malus domestica, Manihot esculenta, Medicago truncatula, Morus notabilis, Musa acuminata, Nelumbo nucifera, Nicotiana sp., Phoenix dactylifera, Phaseolus vulgaris, Populus sp., Prunus mume, Pyrus sp., Ricinus communis, Sesamum indicum, Solanum sp., Spinacia oleracea, Tarenaya hassleriana, Theobroma cacao, Vigna angularis, Vitis vinifera, 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 example

The samples extracted either directly with SDS loading buffer (50mM Tris-HCl pH6.8, 100mM DTT, 2%SDS, 10% glycerol, 0.025% bromophenol blue) and then denatured at 95°C for 5 min, or first extracted with CE buffer (250mM sucrose, 100mM HEPES-KOH pH 7.5, 5% glycerol, 1mM Na₂MoO₄ x 2H₂O, 25mM NaF, 10mM EDTA, 1mM DTT, 0.5%Triton X-100, protease inhibitor cocktail) and then denatured with SDS loading buffer at 70°C for 5 min. 10 µl of proteins were loaded into each well and separated on 10% SDS-PAGE and blotted 1h to PVDF using tank transfer. Blots were blocked with Tris-buffered saline containing 0.05% Tween-20 (TBS-T) and 5% skimmed milk powder for 1h at room temperature (RT) with agitation. The blot was incubated in the primary antibodies indicated at a dilution of 1: 5 000 overnight at 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 5 times for 15 min in TBS-T with milk powder at RT with agitation. Blot was incubated in secondary antibody (Goat-Anti-Rabbit AP conjugate, Sigma) diluted 1:5000 for 2h at RT with agitation. The blot was washed as above with TBS-T without milk powder, equilibrated in AP buffer (100mM TRIS pH=9.5, 100mM NaCl, 50mM MgCl₂) and then developed with chromogenic detection system, before exposure to a CEA RP NEW film for 20 s.

Courtesy of Dr. Elena Petusching, Georg-August-University Goettingen, Germany

Western blot using anti-GPA1 antibodies

Immunodetection of GPA1 (alpha subunit of heterotrimeric G protein). Microsomes were affinity purified to isolate palmitoylated proteins using the acyl biotin exchange method1. Proteins were separated by SDS-PAGE and immunodetected with anti-GPA1. GPA1 is expected to be modified by S-acylation and was enriched in the affinity-purified fraction.

1. Wan, J., Roth, A. F., Bailey, A. O. & Davis, N. G. Palmitoylated proteins: purification and identification. Nat. Protoc. (2007). doi:10.1038/nprot.2007.225

Palmitoylated (S-acylated) proteins were affinity purified from Arabidopsis thaliana using the acyl biotin exchange method1 and separated using SDS-PAGE. Proteins were electro-transferred to PVDF membrane (Millipore, Cat. No. IVPH00010). The membrane was blocked with 5% non-fat dry milk (NFDM) in Tris-buffered saline (TBS) for two hours at room temperature. All incubations were performed with gentle agitation. The membrane was treated with primary antibody (anti-GPA1 [Agrisera, Cat. No. AS12 2370]; 1:7500 in TBS containing 5% NFDM and 0.05% Tween-20) for 2 hours. Blots were quickly rinsed twice with TBS containing 0.05% Tween (TBS-T), then washed five times for 15 minutes each with TBS-T on a shaker. The membrane was treated for 45 minutes with HRP-tagged secondary antibody (Agrisera, Cat. No. AS09 602) diluted 1:10 000 in TBS-T containing 5% non-fat dried milk. Blots were washed 2-3 times for 15 minutes with TBS-T, followed by two 15-minute washes with TBS. Immunolabeled proteins were detected with chemiluminescent detection reagent and imaged using X-ray film.

Courtesy of John McLarney under the guidance of Dr. Estelle Hrabak, University of New Hampshire, USA

Additional information

Background

GPA1 (G PROTEIN ALPHA-1) is involved as a modulator or transducer in various transmembrane signaling systems. Together with GCR1, may regulate the cell cycle via a signaling cascade Promotes abscisic acid (ABA) responses in guard cells. But, together with GCR1 and GB1, acts as a negative regulator of ABA during seed germination and early seedling development. Involved in the blue light (BL) signaling. Alternative name: GP-alpha-1.

Product citations

immunogen:	KLH-conjugated synthetic peptide derived from Arabidopsis thaliana GPA1, UniProt: P18064, TAIR: AT2G26300
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: 2000 (WB)
calculated \| apparent molecular mass [kDa]:	49,3 kDa
Confirmed reactivity:	Arabidopsis thaliana
predicted reactivity:	Amborella trichopoda, Ananas comosus, Arachis sp., Beta vulgaris, Brassica sp., Cajanus cajan, Camelina sativa, Capsella rubella, Capsicum annuum, Cicer arietinum, Coffea canephora, Cucumis melo, Daucus carota, Elaeis guineensis, Eucalyptus grandis, Eutrema sp., Fragaria vesca, Genlisea aurea, Glycine max, Glycine soja, Jatropha curcas, Malus domestica, Manihot esculenta, Medicago truncatula, Morus notabilis, Musa acuminata, Nelumbo nucifera, Nicotiana sp., Phoenix dactylifera, Phaseolus vulgaris, Populus sp., Prunus mume, Pyrus sp., Ricinus communis, Sesamum indicum, Solanum sp., Spinacia oleracea, Tarenaya hassleriana, Theobroma cacao, Vigna angularis, Vitis vinifera, 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 The samples extracted either directly with SDS loading buffer (50mM Tris-HCl pH6.8, 100mM DTT, 2%SDS, 10% glycerol, 0.025% bromophenol blue) and then denatured at 95°C for 5 min, or first extracted with CE buffer (250mM sucrose, 100mM HEPES-KOH pH 7.5, 5% glycerol, 1mM Na₂MoO₄ x 2H₂O, 25mM NaF, 10mM EDTA, 1mM DTT, 0.5%Triton X-100, protease inhibitor cocktail) and then denatured with SDS loading buffer at 70°C for 5 min. 10 µl of proteins were loaded into each well and separated on 10% SDS-PAGE and blotted 1h to PVDF using tank transfer. Blots were blocked with Tris-buffered saline containing 0.05% Tween-20 (TBS-T) and 5% skimmed milk powder for 1h at room temperature (RT) with agitation. The blot was incubated in the primary antibodies indicated at a dilution of 1: 5 000 overnight at 4°C with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed 5 times for 15 min in TBS-T with milk powder at RT with agitation. Blot was incubated in secondary antibody (Goat-Anti-Rabbit AP conjugate, Sigma) diluted 1:5000 for 2h at RT with agitation. The blot was washed as above with TBS-T without milk powder, equilibrated in AP buffer (100mM TRIS pH=9.5, 100mM NaCl, 50mM MgCl₂) and then developed with chromogenic detection system, before exposure to a CEA RP NEW film for 20 s. Courtesy of Dr. Elena Petusching, Georg-August-University Goettingen, Germany Immunodetection of GPA1 (alpha subunit of heterotrimeric G protein). Microsomes were affinity purified to isolate palmitoylated proteins using the acyl biotin exchange method1. Proteins were separated by SDS-PAGE and immunodetected with anti-GPA1. GPA1 is expected to be modified by S-acylation and was enriched in the affinity-purified fraction. 1. Wan, J., Roth, A. F., Bailey, A. O. & Davis, N. G. Palmitoylated proteins: purification and identification. Nat. Protoc. (2007). doi:10.1038/nprot.2007.225 Palmitoylated (S-acylated) proteins were affinity purified from Arabidopsis thaliana using the acyl biotin exchange method1 and separated using SDS-PAGE. Proteins were electro-transferred to PVDF membrane (Millipore, Cat. No. IVPH00010). The membrane was blocked with 5% non-fat dry milk (NFDM) in Tris-buffered saline (TBS) for two hours at room temperature. All incubations were performed with gentle agitation. The membrane was treated with primary antibody (anti-GPA1 [Agrisera, Cat. No. AS12 2370]; 1:7500 in TBS containing 5% NFDM and 0.05% Tween-20) for 2 hours. Blots were quickly rinsed twice with TBS containing 0.05% Tween (TBS-T), then washed five times for 15 minutes each with TBS-T on a shaker. The membrane was treated for 45 minutes with HRP-tagged secondary antibody (Agrisera, Cat. No. AS09 602) diluted 1:10 000 in TBS-T containing 5% non-fat dried milk. Blots were washed 2-3 times for 15 minutes with TBS-T, followed by two 15-minute washes with TBS. Immunolabeled proteins were detected with chemiluminescent detection reagent and imaged using X-ray film. Courtesy of John McLarney under the guidance of Dr. Estelle Hrabak, University of New Hampshire, USA
background:	GPA1 (G PROTEIN ALPHA-1) is involved as a modulator or transducer in various transmembrane signaling systems. Together with GCR1, may regulate the cell cycle via a signaling cascade Promotes abscisic acid (ABA) responses in guard cells. But, together with GCR1 and GB1, acts as a negative regulator of ABA during seed germination and early seedling development. Involved in the blue light (BL) signaling. Alternative name: GP-alpha-1.

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