AtpB | Beta subunit of ATP synthase (chloroplastic + mitochondrial) (chicken antibodies)

Name: AtpB | Beta subunit of ATP synthase (chloroplastic + mitochondrial) (chicken antibodies)
SKU: AS03 030
Price: 394 EUR
Availability: InStock
Rating: 5 (1 reviews)

AS03 030 | Clonality: Polyclonal | Host: Chicken | Reactivity: [global antibody] for plant and bacterial F-type ATP synthases

Benefits of using this antibody

AtpB | Beta subunit of ATP synthase (chloroplastic + mitochondrial) (chicken antibodies) in the group Antibodies Plant/Algal / Global Antibodies at Agrisera AB (Antibodies for research) (AS03 030)

AtpB | Beta subunit of ATP synthase (chloroplastic + mitochondrial) (chicken antibodies)

Product Information

Immunogen

KLH-conjugated synthetic peptide derived from available plant, algal (chloroplastic and mitochondrial) and bacterial sequences of beta subunits of F-type ATP synthases, including Arabidopsis thaliana chloroplastic ATP synthase subunit beta UniProt: P19366, TAIR: AtCg00480 and Arabidopsis thaliana mitochondrial ATP synthase subunit beta-1, UniProt: P83483, TAIR: At5g08670 as well as Chlamydomonas reinhardtii, UniProt: P06541 and A8IQU3

Host Chicken

Clonality Polyclonal

Purity Purified, total IgY (chicken egg yolk immunoglobulin) in PBS pH 8. Contains 0.02 % sodium azide.

Format Liquid at 23,3 µg/µl

Quantity 100 µl

Storage Store at 2-8°C.; make aliquots to avoid working with a stock. 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 Immunolocalization (IL), Immunogold (IG), Western blot (WB)

Recommended dilution 1 : 500 for localization of native enzyme by immunogold (IL), 1 : 5 000-1 : 8 000 (WB)

Expected | apparent MW

53.9 kDa (Arabidopsis thaliana), 51.7 kDa (Synechocystis PCC 6803), 53.7 kDa (Spinacia oleracea)

Reactivity

Confirmed reactivity Arabidopsis thaliana, Hordeum vulgare, Phaeodactylum tricornutum, Spartina alterniflora, Spinacia oleracea, Synechocystis PCC 6803, Synechococcus PCC 7942, beef muscle, Rat liver

Predicted reactivity Acinetobacter baumannii, Bacillus subtilis, Brassica napus, Cannabis sativa, Clostridioides difficile, Cyanobacteria, E.coli K-12, Galdieria sulphuraria, Heliomicrobium modesticaldum Ice1, Manihot esculenta, Nicotiana plumbaginifolia, Saccharomyces cerevisiae, Salmonella typhimurium, Trichodesmium erythraeum, Triticum aestivum, Vitis vinifera, Zosteria marina, Yrsinia sp.

Species of your interest not listed? Contact us

Not reactive in Archeal V-type ATP synthase

Application examples

Application examples application information

10 µg of total protein from samples such as beef muscle (1), rat liver (2), Arabidopsis thaliana leaf (3), Hordeum vulgare leaf (4), Synechocystis PCC 6803 total cell (5), Synechococcus PCC 7942 total cell (6) were extracted with Protein Extraction Buffer PEB (AS08 300). Samples were diluted with 1X sample buffer (NuPAGE LDS sample buffer (Invitrogen) supplemented with 50 mM DTT and heat at 70C for 5 min and keep on ice before loading. Protein samples were separated on Bolt 4-12% Tris-Bis gel (Invitrogen) LDS-PAGE and blotted for 1h to 1.5h on PVDF using Bolt Mini Blot Module. Blot was blocked immediately following transfer in 2% blocking reagent (GE RPN 2125; Healthcare) dissolved in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5 000 (in blocking reagent) for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, and then washed 1x15 min and 3x5 min with TBS-T at room temperature with agitation. Blot was incubated in secondary antibody (anti-chicken IgY horse radish peroxidase conjugated) diluted to 1:20 000 in blocking reagent for 1h at room temperature with agitation. The blot was washed as above. Signals in the blot were detected using Lumigen ECL Ultra Reagent (Lumigen TMA-6, Lumigen), and visualized using the Molecular Imager VersaDoc MP 4000 System (Bio-Rad) and Quantity One software (Bio-Rad). Exposure time was 30 seconds.

Reactant: Chlamydomonas reinhardtii (Green Alga)

Application: Western Blotting

Pudmed ID: 25653846

Journal: F1000Res

Figure Number: 1A

Published Date: 2015-02-06

First Author: Johnson, E. A. & Lecomte, J. T.

Impact Factor: None

Open Publication

Purification of THB1 fromC. reinhardtii cell culture.Protein samples from different stages of purification were analyzed by electrophoresis. (A) Samples of protein extracts before and after lysis ofC. reinhardtii cells with liquid nitrogen. Proteins separated on 16.5% Tris-tricine gel then transferred to nitrocellulose and immunostained with antibodies against proteins localized to different cellular compartments. Lysis by liquid nitrogen enriches the lysate with soluble proteins without enrichment of proteins found in major algal organelles. Histone H3 protein is located in the nucleus, nitrate reductase is a soluble cytosolic protein and the ATP synthase ? subunit is part of the thylakoid membrane of the chloroplast. (B) Samples from different steps in the purification procedure were separated by electrophoresis. Following separation, the proteins within the gel were visualized using silver stain. (C) Proteins prepared identically to those detected in panel B were transferred to nitrocellulose followed by immunostaining with polyclonal antibodies specific for THB1. In addition to the protein samples, a lane was used for molecular weight markers (Spectra LR, ThermoScientific). The numbers indicated between the panels represents location of the markers (kDa molecular mass). WC, whole cell protein extract. Lys, protein extract lysate, and Pel, protein pellet following liquid nitrogen fracturing. QFF, concentrated sample following anion exchange chromatography. S75, concentrated sample following separation on the Superdex 75 column.

Additional information

Additional information The anti-AtpB antibody will detect the mitochondrial form of the F1 ATP synthase subcomplex, as well as the chloroplastic CF1 ATP synthase and most known bacterial F-type ATP synthases, Peptide used for antibody production is located in a beta sheet, which is partly exposed near the surface of the AtpB protein

Results of immunogold studies using anti-AtpB antibody are published in Andersson et al. (2009).

Background

ATP synthase is the universal enzyme that synthesizes ATP from ADP and phosphate using the energy stored in a transmembrane ion gradient.

Product citations

Selected references Neusius et al. (2022) Lysine acetylation regulates moonlighting activity of the E2 subunit of the chloroplast pyruvate dehydrogenase complex in Chlamydomonas. Plant J. 2022 Sep;111(6):1780-1800. doi: 10.1111/tpj.15924. Epub 2022 Aug 8. PMID: 35899410.
Levitan et al. (2019). Structural and functional analyses of photosystem II in the marine diatom Phaeodactylum tricornutum. Proc Natl Acad Sci U S A. 2019 Aug 27;116(35):17316-17322. doi: 10.1073/pnas.1906726116.
Nelson et al. (2019). Protein lysine methylation contributes to modulating the response of sensitive and tolerant Arabidopsis species to cadmium stress. doi: 10.1111/pce.13692.
Gellert et al. (2018). A single point mutation on the cucumber mosaic virus surface induces an unexpected and strong interaction with the F1 complex of the ATP synthase in Nicotiana clevelandii plants. Virus Res. 2018 Jun 2;251:47-55. doi: 10.1016/j.virusres.2018.05.005. (immunogold)
Quesada et al. (2011). Arabidopsis RUGOSA2 encodes an mTERF family member required for mitochondrion, chloroplast and leaf development. Plant J. Nov;68(4):738-53. doi: 10.1111/j.1365-313X.2011.04726.x. Epub 2011 Sep 13.
Andersson et. al (2009). Co-localization of P-glycerate kinase, P-ribulokinase, ADP-glucose pyrophosphorylase and Rubisco activase with CF1 in pea leaf chloroplasts. Plant Science 177:136-14. (immunologold)
Morash et al. (2007). Macromolecular dynamics of the photosynthetic system over a seasonal developmental progression in Spartina alterniflora. Canadian J. of Botany, 2007, 85(5): 476-483, 10.1139/B07-043.

immunogen:	KLH-conjugated synthetic peptide derived from available plant, algal (chloroplastic and mitochondrial) and bacterial sequences of beta subunits of F-type ATP synthases, including Arabidopsis thaliana chloroplastic ATP synthase subunit beta UniProt: P19366, TAIR: AtCg00480 and Arabidopsis thaliana mitochondrial ATP synthase subunit beta-1, UniProt: P83483, TAIR: At5g08670 as well as Chlamydomonas reinhardtii, UniProt: P06541 and A8IQU3
Host:	Chicken
Clonality:	Polyclonal
Purity:	Purified, total IgY (chicken egg yolk immunoglobulin) in PBS pH 8. Contains 0.02 % sodium azide.
Format:	Liquid at 23,3 µg/µl
Quantity:	100 µl
storage:	Store at 2-8°C.; make aliquots to avoid working with a stock. 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:	Immunolocalization (IL), Immunogold (IG), Western blot (WB)
recommended dilution:	1 : 500 for localization of native enzyme by immunogold (IL), 1 : 5 000-1 : 8 000 (WB)
Expected \| apparent MW:	53.9 kDa (Arabidopsis thaliana), 51.7 kDa (Synechocystis PCC 6803), 53.7 kDa (Spinacia oleracea)
Confirmed reactivity:	Arabidopsis thaliana, Hordeum vulgare, Phaeodactylum tricornutum, Spartina alterniflora, Spinacia oleracea, Synechocystis PCC 6803, Synechococcus PCC 7942, beef muscle, Rat liver
predicted reactivity:	Acinetobacter baumannii, Bacillus subtilis, Brassica napus, Cannabis sativa, Clostridioides difficile, Cyanobacteria, E.coli K-12, Galdieria sulphuraria, Heliomicrobium modesticaldum Ice1, Manihot esculenta, Nicotiana plumbaginifolia, Saccharomyces cerevisiae, Salmonella typhimurium, Trichodesmium erythraeum, Triticum aestivum, Vitis vinifera, Zosteria marina, Yrsinia sp. Species of your interest not listed? Contact us
not reactive in:	Archeal V-type ATP synthase
Picture (footer):	application information 10 µg of total protein from samples such as beef muscle (1), rat liver (2), Arabidopsis thaliana leaf (3), Hordeum vulgare leaf (4), Synechocystis PCC 6803 total cell (5), Synechococcus PCC 7942 total cell (6) were extracted with Protein Extraction Buffer PEB (AS08 300). Samples were diluted with 1X sample buffer (NuPAGE LDS sample buffer (Invitrogen) supplemented with 50 mM DTT and heat at 70C for 5 min and keep on ice before loading. Protein samples were separated on Bolt 4-12% Tris-Bis gel (Invitrogen) LDS-PAGE and blotted for 1h to 1.5h on PVDF using Bolt Mini Blot Module. Blot was blocked immediately following transfer in 2% blocking reagent (GE RPN 2125; Healthcare) dissolved in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5 000 (in blocking reagent) for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, and then washed 1x15 min and 3x5 min with TBS-T at room temperature with agitation. Blot was incubated in secondary antibody (anti-chicken IgY horse radish peroxidase conjugated) diluted to 1:20 000 in blocking reagent for 1h at room temperature with agitation. The blot was washed as above. Signals in the blot were detected using Lumigen ECL Ultra Reagent (Lumigen TMA-6, Lumigen), and visualized using the Molecular Imager VersaDoc MP 4000 System (Bio-Rad) and Quantity One software (Bio-Rad). Exposure time was 30 seconds.
More images:	Reactant: Chlamydomonas reinhardtii (Green Alga) Application: Western Blotting Pudmed ID: 25653846 Journal: F1000Res Figure Number: 1A Published Date: 2015-02-06 First Author: Johnson, E. A. & Lecomte, J. T. Impact Factor: None Open Publication Purification of THB1 fromC. reinhardtii cell culture.Protein samples from different stages of purification were analyzed by electrophoresis. (A) Samples of protein extracts before and after lysis ofC. reinhardtii cells with liquid nitrogen. Proteins separated on 16.5% Tris-tricine gel then transferred to nitrocellulose and immunostained with antibodies against proteins localized to different cellular compartments. Lysis by liquid nitrogen enriches the lysate with soluble proteins without enrichment of proteins found in major algal organelles. Histone H3 protein is located in the nucleus, nitrate reductase is a soluble cytosolic protein and the ATP synthase ? subunit is part of the thylakoid membrane of the chloroplast. (B) Samples from different steps in the purification procedure were separated by electrophoresis. Following separation, the proteins within the gel were visualized using silver stain. (C) Proteins prepared identically to those detected in panel B were transferred to nitrocellulose followed by immunostaining with polyclonal antibodies specific for THB1. In addition to the protein samples, a lane was used for molecular weight markers (Spectra LR, ThermoScientific). The numbers indicated between the panels represents location of the markers (kDa molecular mass). WC, whole cell protein extract. Lys, protein extract lysate, and Pel, protein pellet following liquid nitrogen fracturing. QFF, concentrated sample following anion exchange chromatography. S75, concentrated sample following separation on the Superdex 75 column.
additional information:	The anti-AtpB antibody will detect the mitochondrial form of the F1 ATP synthase subcomplex, as well as the chloroplastic CF1 ATP synthase and most known bacterial F-type ATP synthases, Peptide used for antibody production is located in a beta sheet, which is partly exposed near the surface of the AtpB protein
additional information (application):	Results of immunogold studies using anti-AtpB antibody are published in Andersson et al. (2009).
background:	ATP synthase is the universal enzyme that synthesizes ATP from ADP and phosphate using the energy stored in a transmembrane ion gradient.
All references:	Neusius et al. (2022) Lysine acetylation regulates moonlighting activity of the E2 subunit of the chloroplast pyruvate dehydrogenase complex in Chlamydomonas. Plant J. 2022 Sep;111(6):1780-1800. doi: 10.1111/tpj.15924. Epub 2022 Aug 8. PMID: 35899410. Levitan et al. (2019). Structural and functional analyses of photosystem II in the marine diatom Phaeodactylum tricornutum. Proc Natl Acad Sci U S A. 2019 Aug 27;116(35):17316-17322. doi: 10.1073/pnas.1906726116. Nelson et al. (2019). Protein lysine methylation contributes to modulating the response of sensitive and tolerant Arabidopsis species to cadmium stress. doi: 10.1111/pce.13692. Gellert et al. (2018). A single point mutation on the cucumber mosaic virus surface induces an unexpected and strong interaction with the F1 complex of the ATP synthase in Nicotiana clevelandii plants. Virus Res. 2018 Jun 2;251:47-55. doi: 10.1016/j.virusres.2018.05.005. (immunogold) Quesada et al. (2011). Arabidopsis RUGOSA2 encodes an mTERF family member required for mitochondrion, chloroplast and leaf development. Plant J. Nov;68(4):738-53. doi: 10.1111/j.1365-313X.2011.04726.x. Epub 2011 Sep 13. Andersson et. al (2009). Co-localization of P-glycerate kinase, P-ribulokinase, ADP-glucose pyrophosphorylase and Rubisco activase with CF1 in pea leaf chloroplasts. Plant Science 177:136-14. (immunologold) Morash et al. (2007). Macromolecular dynamics of the photosynthetic system over a seasonal developmental progression in Spartina alterniflora. Canadian J. of Botany, 2007, 85(5): 476-483, 10.1139/B07-043.