Goat anti-Rabbit IgG (H&L), HRP conjugated

AS09 602 | Clonality: Polyclonal | Host: Goat | Reactivity: Rabbit IgG (H&L)

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Goat anti-Rabbit IgG (H&L), HRP conjugated

Product Information

Immunogen Purified Rabbit IgG, whole molecule,

Host Goat

Clonality Polyclonal

Purity Immunogen affinity purified using solid phase rabbit IgG.

Format Lyophilized

Quantity 1 mg

Reconstitution For reconstitution add 1,1 ml of sterile water, Let it stand 30 minutes at room temperature to dissolve, Spin centrifuge shortly to remove any particles, Prepare fresh working dilutions daily

Storage Store lyophilized material at 2-8°C. For long time storage after reconstitution, dilute the antibody solution with glycerol to a final concentration of 50% glycerol and store as liquid at -20°C, to prevent loss of enzymatic activity. For example, if you have reconstituted 1 mg of antibody in 1,1 ml of sterile water add 1,1 ml of glycerol. Such solution will not freeze in -20°C. If you are using a 1:5000 dilution prior to diluting with glycerol, then you would need to use a 1:2500 dilution after adding glycerol. Prepare working dilution prior to use and then discard, Be sure to mix well but without foaming.

Tested applications ELISA (ELISA), Immunohistochemistry (IHC), Western blot (WB)

Recommended dilution 1 : 50 000 -1 : 90 000 (ELISA), 1 : 500 -1 : 5000 (IHC), 1: 10 000 -1 : 50 000 (WB)

Reactivity

Confirmed reactivity Based on IEP, this antibody reacts with: rabbit IgG heavy chains and light chains on all rabbit immunoglobulins

Not reactive in Non-immunoglobulin rabbit serum proteins

Application examples

Application examples Application example

western blot detection of PsaC using Agrisera primary and secondary antibodies

5 µg of total extract from (1) Hordeum vulgaretotal leaf, (2) Zea mays (3) Spinacia oleracea extracted with PEB (AS08 300) were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% ECL Advance blocking reagent (GE Healthcare) 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. Blots were incubated in the primary anti-PsaC antibody (AS04 042) at a dilution of 1: 10 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (goat anti-rabbit IgG horse radish peroxidase conjugated, AS09 602, Agrisera) diluted to 1:50 000 in 2% ECL Advance blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with chemiluminescent detection reagent according to the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad). Exposure time was 30 seconds.

Comparison of Agrisera secondary antibody sensitivity

comparison of secondary antibody sensitivity

10 μg of mitochondrial fraction from Arabidopsis thaliana (1,3) and Arabidopsis thaliana leaf extract (2,4) were separated on 10% gel and blotted on nitrocellulose membrane using wet transfer (0.22% CAPS, pH 11). Filters where blocked (1.5h) in 5% milk in TBST (1X TBS, 0,1% Tween 20), incubated with 1: 1000 anti-COXII antibodies (2h in TBST) followed by incubation with 1: 10 000 secondary anti-rabbit (1h) HRP-coupled antibodies from Agrisera (left panel) and other manufacture (right panel) and visualized with chemiluminescent detection reagent, on Kodak autoradiography film for 5 s. Antibody in left panel detects target protein also in total cell extract (2) and can be used in higher dilution than applied 1: 10 000.

Agrisera goat anti-rabbit HRP conjugated antibody (AS09 602) can be used at following dilutions: 1: 50 000 -1: 90 000 (ELISA), 1 : 75 000 with chemiluminescence detection range of extreme low picogram and 1: 25 000 with chemiluminescence detection reagent of mid femtogram (WB), 1: 500 -1: 5000 (IHC).

Additional information

Concentration: 1.0 mg/ml.

Antibody is provided in: 10 mM Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 1% BSA (w/v), Protease IgG free, 0.1 % (v/v) ProClin 150.

Affinity purified antibody is > 95 % pure, according to SDS-PAGE.

This antibody can used on a very wide range of samples from various species including many model plants, algae, diatoms and bacteria.

Background

Goat anti-rabbit IgG (H&L) is a secondary antibody conjugated to HRP which binds to all rabbit IgGs in immunological assays.

Product citations

Selected references Lim et al (2022). Arabidopsis guard cell chloroplasts import cytosolic ATP for starch turnover and stomatal opening. Nat Commun. 2022 Feb 3;13(1):652. doi: 10.1038/s41467-022-28263-2. PMID: 35115512; PMCID: PMC8814037.
Miklankova et al. (2022) HYPK promotes the activity of the Nalfa-acetyltransferase A complex to determine proteostasis of nonAc-X2/N-degron-containing proteins. Sci Adv. 2022 Jun 17;8(24):eabn6153. doi: 10.1126/sciadv.abn6153. Epub 2022 Jun 15. PMID: 35704578; PMCID: PMC9200280.
Hofmann, Wienkoop & Luthje (2022) Hypoxia-Induced Aquaporins and Regulation of Redox Homeostasis by a Trans-Plasma Membrane Electron Transport System in Maize Roots. Antioxidants (Basel). 2022 Apr 25;11(5):836. doi: 10.3390/antiox11050836. PMID: 35624700; PMCID: PMC9137787.
Bychkov et al. (2022) The role of PAP4/FSD3 and PAP9/FSD2 in heat stress responses of chloroplast genes. Plant Sci. 2022 Sep;322:111359. doi: 10.1016/j.plantsci.2022.111359. Epub 2022 Jun 20. PMID: 35738478.
Vitale et al. (2021) Light Spectral Composition Influences Structural and Eco-Physiological Traits of Solanum lycopersicum L. cv. 'Microtom' in Response to High-LET Ionizing Radiation. Plants (Basel). 2021 Aug 23;10(8):1752. doi: 10.3390/plants10081752. PMID: 34451797; PMCID: PMC8399554.
Gucum et al. (2021) A patient-based medaka alg2 mutant as a model for hypo-N-glycosylation. Development. 2021 Jun 1;148(11):dev199385. doi: 10.1242/dev.199385. Epub 2021 Jun 9. PMID: 34106226; PMCID: PMC8217707.
Wu et al. (2021). Formation of light-harvesting complex (LHC) II aggregates from LHCII-PSI-LHCI complexes in rice plants under high light. J Exp Bot. 2021 May 3:erab188. doi: 10.1093/jxb/erab188. Epub ahead of print. PMID: 33939808.
Wojciechowska et al. (2021) Localization and Dynamics of the Methionine Sulfoxide Reductases MsrB1 and MsrB2 in Beech Seeds. Int J Mol Sci. 2021 Jan 2;22(1):402. doi: 10.3390/ijms22010402. PMID: 33401671; PMCID: PMC7795007.
Long et al. (2020). FOXO3 is targeted by miR-223-3p and promotes osteogenic differentiation of bone marrow mesenchymal stem cells by enhancing autophagy. Hum Cell. 2020 Sep 13.doi: 10.1007/s13577-020-00421-y.
Khajuria et al. (2020). Photochemical Efficiency Is Negatively Correlated With the Delta 9- Tetrahydrocannabinol Content in Cannabis Sativa L. Plant Physiol Biochem. 2020 Apr 8;151:589-600. doi: 10.1016/j.plaphy.2020.04.003.
Ignatov et al. (2020). An mRNA-mRNA Interaction Couples Expression of a Virulence Factor and Its Chaperone in Listeria Monocytogenes. Cell Rep, 30 (12), 4027-4040.e7
Hu et al. (2019). Photoprotection capacity of microalgae improved by regulating the antenna size of light-harvesting complexes. J Appl Phycol (2019). doi.org/10.1007/s10811-019-01969-5
Schober et al. (2019). Organelle Studies and Proteome Analyses on Mitochondria and Plastids Fractions from the Diatom Thalassiosira pseudonana. Plant Cell Physiol. 2019 Jun 10. pii: pcz097. doi: 10.1093/pcp/pcz097.
Rethore et al. (2019). Arabidopsis seedlings display a remarkable resilience under severe mineral starvation using their metabolic plasticity to remain self-sufficient for weeks. Plant J. 2019 Mar 22. doi: 10.1111/tpj.14325.
Contreras et al. (2019). UV-B shock induces photoprotective flavonoids but not antioxidant activity in Antarctic Colobanthus quitensis (Kunth) Bartl. Environmental and Experimental Botany, Volume 159, March 2019, Pages 179-190.
Nikkanen et al. (2018). Multilevel regulation of non-photochemical quenching and state transitions by chloroplast NADPH-dependent thioredoxin reductase. Physiol Plant. 2018 Dec 22. doi: 10.1111/ppl.12914.
Migocka et al. (2018). Cucumber metal tolerance protein 7 (CsMTP7) is involved in the accumulation of Fe in mitochondria under Fe excess. Plant J. 2018 Jun 22. doi: 10.1111/tpj.14006.
Tong et al. (2018). Delivery of siRNA in vitro and in vivo using PEI-capped porous silicon nanoparticles to silence MRP1 and inhibit proliferation in glioblastoma. J Nanobiotechnology. 2018 Apr 13;16(1):38. doi: 10.1186/s12951-018-0365-y.
Nikkanen et al. (2018). Regulation of chloroplast NADH dehydrogenase-like complex by NADPH-dependent thioredoxin system. CSH, BioRixiv. doi.org/10.1101/261560.
Sinclair et al. (2017) Etiolated Seedling Development Requires Repression of Photomorphogenesis by a Small Cell-Wall-Derived Dark Signal. Curr Biol. 2017 Nov 20;27(22):3403-3418.e7. doi: 10.1016/j.cub.2017.09.063.
Gzyl et al. (2017). Gamma-tubulin distribution and ultrastructural changes in root cells of soybean (Glycine max L.) seedlings under cadmium stress. Environmental and Experimental Botany, Vol 143, Nov 2017, Pages 82-90.
Kamies et al. (2017). A Proteomic Approach to Investigate the Drought Response in the Orphan Crop Eragrostis tef. Proteomes. 2017 Nov 15;5(4). pii: E32. doi: 10.3390/proteomes5040032.
Niederhuber et al. (2017). Super-resolution microscopy of the ß-carboxysome reveals a homogenous matrix. Mol Biol Cell. 2017 Aug 9. pii: mbc.E17-01-0069. doi: 10.1091/mbc.E17-01-0069.
Tyuereva et al. (2017). The absence of chlorophyll b affects lateral mobility of photosynthetic complexes and lipids in grana membranes of Arabidopsis and barley chlorina mutants. Photosynth Res. 2017 Apr 5. doi: 10.1007/s11120-017-0376-9. (Hordeum vulgare, western blot)
Romanowska et al. (2017). Differences in photosynthetic responses of NADP-ME type C4 species to high light. Planta. 2017 Mar;245(3):641-657. doi: 10.1007/s00425-016-2632-1. Epub 2016 Dec 18.
Good et al. (2016). Attenuating Listeria monocytogenes Virulence by Targeting the Regulatory Protein PrfA. Cell Chem Biol. 2016 Mar 17;23(3):404-14. doi: 10.1016/j.chembiol.2016.02.013.
Datta et al. (2016). Glutathione Regulates 1-Aminocyclopropane-1-Carboxylate Synthase Transcription via WRKY33 and 1-Aminocyclopropane-1-Carboxylate Oxidase by Modulating Messenger RNA Stability to Induce Ethylene Synthesis during Stress. Plant Physiol. 2015 Dec;169(4):2963-81. doi: 10.1104/pp.15.01543. Epub 2015 Oct 13.
Barahimipour et al. (2016). Efficient expression of nuclear transgenes in the green alga Chlamydomonas: synthesis of an HIV antigen and development of a new selectable marker. Plant Mol Biol. 2016 Mar;90(4-5):403-18. doi: 10.1007/s11103-015-0425-8. Epub 2016 Jan 8.
CiteAB has listed this antibody as one of the 7000 most published antibodies in the world.

Picture (footer):	Application example 5 µg of total extract from (1) Hordeum vulgaretotal leaf, (2) Zea mays (3) Spinacia oleracea extracted with PEB (AS08 300) were separated on 4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% ECL Advance blocking reagent (GE Healthcare) 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. Blots were incubated in the primary anti-PsaC antibody (AS04 042) at a dilution of 1: 10 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (goat anti-rabbit IgG horse radish peroxidase conjugated, AS09 602, Agrisera) diluted to 1:50 000 in 2% ECL Advance blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with chemiluminescent detection reagent according to the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad). Exposure time was 30 seconds. Comparison of Agrisera secondary antibody sensitivity 10 μg of mitochondrial fraction from Arabidopsis thaliana (1,3) and Arabidopsis thaliana leaf extract (2,4) were separated on 10% gel and blotted on nitrocellulose membrane using wet transfer (0.22% CAPS, pH 11). Filters where blocked (1.5h) in 5% milk in TBST (1X TBS, 0,1% Tween 20), incubated with 1: 1000 anti-COXII antibodies (2h in TBST) followed by incubation with 1: 10 000 secondary anti-rabbit (1h) HRP-coupled antibodies from Agrisera (left panel) and other manufacture (right panel) and visualized with chemiluminescent detection reagent, on Kodak autoradiography film for 5 s. Antibody in left panel detects target protein also in total cell extract (2) and can be used in higher dilution than applied 1: 10 000. Agrisera goat anti-rabbit HRP conjugated antibody (AS09 602) can be used at following dilutions: 1: 50 000 -1: 90 000 (ELISA), 1 : 75 000 with chemiluminescence detection range of extreme low picogram and 1: 25 000 with chemiluminescence detection reagent of mid femtogram (WB), 1: 500 -1: 5000 (IHC).
Clonality:	Polyclonal
Format:	Lyophilized
Host:	Goat
immunogen:	Purified Rabbit IgG, whole molecule,
Purity:	Immunogen affinity purified using solid phase rabbit IgG.
Quantity:	1 mg
recommended dilution:	1 : 50 000 -1 : 90 000 (ELISA), 1 : 500 -1 : 5000 (IHC), 1: 10 000 -1 : 50 000 (WB)
Reconstitution:	For reconstitution add 1,1 ml of sterile water, Let it stand 30 minutes at room temperature to dissolve, Spin centrifuge shortly to remove any particles, Prepare fresh working dilutions daily
storage:	Store lyophilized material at 2-8°C. For long time storage after reconstitution, dilute the antibody solution with glycerol to a final concentration of 50% glycerol and store as liquid at -20°C, to prevent loss of enzymatic activity. For example, if you have reconstituted 1 mg of antibody in 1,1 ml of sterile water add 1,1 ml of glycerol. Such solution will not freeze in -20°C. If you are using a 1:5000 dilution prior to diluting with glycerol, then you would need to use a 1:2500 dilution after adding glycerol. Prepare working dilution prior to use and then discard, Be sure to mix well but without foaming.
tested applications:	ELISA (ELISA), Immunohistochemistry (IHC), Western blot (WB)
additional information:	Concentration: 1.0 mg/ml. Antibody is provided in: 10 mM Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 1% BSA (w/v), Protease IgG free, 0.1 % (v/v) ProClin 150. Affinity purified antibody is > 95 % pure, according to SDS-PAGE. This antibody can used on a very wide range of samples from various species including many model plants, algae, diatoms and bacteria.
Confirmed reactivity:	Based on IEP, this antibody reacts with: rabbit IgG heavy chains and light chains on all rabbit immunoglobulins
not reactive in:	Non-immunoglobulin rabbit serum proteins
background:	Goat anti-rabbit IgG (H&L) is a secondary antibody conjugated to HRP which binds to all rabbit IgGs in immunological assays.
All references:	Lim et al (2022). Arabidopsis guard cell chloroplasts import cytosolic ATP for starch turnover and stomatal opening. Nat Commun. 2022 Feb 3;13(1):652. doi: 10.1038/s41467-022-28263-2. PMID: 35115512; PMCID: PMC8814037. Miklankova et al. (2022) HYPK promotes the activity of the Nalfa-acetyltransferase A complex to determine proteostasis of nonAc-X2/N-degron-containing proteins. Sci Adv. 2022 Jun 17;8(24):eabn6153. doi: 10.1126/sciadv.abn6153. Epub 2022 Jun 15. PMID: 35704578; PMCID: PMC9200280. Hofmann, Wienkoop & Luthje (2022) Hypoxia-Induced Aquaporins and Regulation of Redox Homeostasis by a Trans-Plasma Membrane Electron Transport System in Maize Roots. Antioxidants (Basel). 2022 Apr 25;11(5):836. doi: 10.3390/antiox11050836. PMID: 35624700; PMCID: PMC9137787. Bychkov et al. (2022) The role of PAP4/FSD3 and PAP9/FSD2 in heat stress responses of chloroplast genes. Plant Sci. 2022 Sep;322:111359. doi: 10.1016/j.plantsci.2022.111359. Epub 2022 Jun 20. PMID: 35738478. Vitale et al. (2021) Light Spectral Composition Influences Structural and Eco-Physiological Traits of Solanum lycopersicum L. cv. 'Microtom' in Response to High-LET Ionizing Radiation. Plants (Basel). 2021 Aug 23;10(8):1752. doi: 10.3390/plants10081752. PMID: 34451797; PMCID: PMC8399554. Gucum et al. (2021) A patient-based medaka alg2 mutant as a model for hypo-N-glycosylation. Development. 2021 Jun 1;148(11):dev199385. doi: 10.1242/dev.199385. Epub 2021 Jun 9. PMID: 34106226; PMCID: PMC8217707. Wu et al. (2021). Formation of light-harvesting complex (LHC) II aggregates from LHCII-PSI-LHCI complexes in rice plants under high light. J Exp Bot. 2021 May 3:erab188. doi: 10.1093/jxb/erab188. Epub ahead of print. PMID: 33939808. Wojciechowska et al. (2021) Localization and Dynamics of the Methionine Sulfoxide Reductases MsrB1 and MsrB2 in Beech Seeds. Int J Mol Sci. 2021 Jan 2;22(1):402. doi: 10.3390/ijms22010402. PMID: 33401671; PMCID: PMC7795007. Long et al. (2020). FOXO3 is targeted by miR-223-3p and promotes osteogenic differentiation of bone marrow mesenchymal stem cells by enhancing autophagy. Hum Cell. 2020 Sep 13.doi: 10.1007/s13577-020-00421-y. Khajuria et al. (2020). Photochemical Efficiency Is Negatively Correlated With the Delta 9- Tetrahydrocannabinol Content in Cannabis Sativa L. Plant Physiol Biochem. 2020 Apr 8;151:589-600. doi: 10.1016/j.plaphy.2020.04.003. Ignatov et al. (2020). An mRNA-mRNA Interaction Couples Expression of a Virulence Factor and Its Chaperone in Listeria Monocytogenes. Cell Rep, 30 (12), 4027-4040.e7 Hu et al. (2019). Photoprotection capacity of microalgae improved by regulating the antenna size of light-harvesting complexes. J Appl Phycol (2019). doi.org/10.1007/s10811-019-01969-5 Schober et al. (2019). Organelle Studies and Proteome Analyses on Mitochondria and Plastids Fractions from the Diatom Thalassiosira pseudonana. Plant Cell Physiol. 2019 Jun 10. pii: pcz097. doi: 10.1093/pcp/pcz097. Rethore et al. (2019). Arabidopsis seedlings display a remarkable resilience under severe mineral starvation using their metabolic plasticity to remain self-sufficient for weeks. Plant J. 2019 Mar 22. doi: 10.1111/tpj.14325. Contreras et al. (2019). UV-B shock induces photoprotective flavonoids but not antioxidant activity in Antarctic Colobanthus quitensis (Kunth) Bartl. Environmental and Experimental Botany, Volume 159, March 2019, Pages 179-190. Nikkanen et al. (2018). Multilevel regulation of non-photochemical quenching and state transitions by chloroplast NADPH-dependent thioredoxin reductase. Physiol Plant. 2018 Dec 22. doi: 10.1111/ppl.12914. Migocka et al. (2018). Cucumber metal tolerance protein 7 (CsMTP7) is involved in the accumulation of Fe in mitochondria under Fe excess. Plant J. 2018 Jun 22. doi: 10.1111/tpj.14006. Tong et al. (2018). Delivery of siRNA in vitro and in vivo using PEI-capped porous silicon nanoparticles to silence MRP1 and inhibit proliferation in glioblastoma. J Nanobiotechnology. 2018 Apr 13;16(1):38. doi: 10.1186/s12951-018-0365-y. Nikkanen et al. (2018). Regulation of chloroplast NADH dehydrogenase-like complex by NADPH-dependent thioredoxin system. CSH, BioRixiv. doi.org/10.1101/261560. Sinclair et al. (2017) Etiolated Seedling Development Requires Repression of Photomorphogenesis by a Small Cell-Wall-Derived Dark Signal. Curr Biol. 2017 Nov 20;27(22):3403-3418.e7. doi: 10.1016/j.cub.2017.09.063. Gzyl et al. (2017). Gamma-tubulin distribution and ultrastructural changes in root cells of soybean (Glycine max L.) seedlings under cadmium stress. Environmental and Experimental Botany, Vol 143, Nov 2017, Pages 82-90. Kamies et al. (2017). A Proteomic Approach to Investigate the Drought Response in the Orphan Crop Eragrostis tef. Proteomes. 2017 Nov 15;5(4). pii: E32. doi: 10.3390/proteomes5040032. Niederhuber et al. (2017). Super-resolution microscopy of the ß-carboxysome reveals a homogenous matrix. Mol Biol Cell. 2017 Aug 9. pii: mbc.E17-01-0069. doi: 10.1091/mbc.E17-01-0069. Tyuereva et al. (2017). The absence of chlorophyll b affects lateral mobility of photosynthetic complexes and lipids in grana membranes of Arabidopsis and barley chlorina mutants. Photosynth Res. 2017 Apr 5. doi: 10.1007/s11120-017-0376-9. (Hordeum vulgare, western blot) Romanowska et al. (2017). Differences in photosynthetic responses of NADP-ME type C4 species to high light. Planta. 2017 Mar;245(3):641-657. doi: 10.1007/s00425-016-2632-1. Epub 2016 Dec 18. Good et al. (2016). Attenuating Listeria monocytogenes Virulence by Targeting the Regulatory Protein PrfA. Cell Chem Biol. 2016 Mar 17;23(3):404-14. doi: 10.1016/j.chembiol.2016.02.013. Datta et al. (2016). Glutathione Regulates 1-Aminocyclopropane-1-Carboxylate Synthase Transcription via WRKY33 and 1-Aminocyclopropane-1-Carboxylate Oxidase by Modulating Messenger RNA Stability to Induce Ethylene Synthesis during Stress. Plant Physiol. 2015 Dec;169(4):2963-81. doi: 10.1104/pp.15.01543. Epub 2015 Oct 13. Barahimipour et al. (2016). Efficient expression of nuclear transgenes in the green alga Chlamydomonas: synthesis of an HIV antigen and development of a new selectable marker. Plant Mol Biol. 2016 Mar;90(4-5):403-18. doi: 10.1007/s11103-015-0425-8. Epub 2016 Jan 8. CiteAB has listed this antibody as one of the 7000 most published antibodies in the world.