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

Product no: AS09 602

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

213
Add to cart
Delivery:  3-6 business days
  • Product Info
  • 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 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
  • 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
  • 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: Liu et al. (2023). RBPome identification in egg-cell like callus of Arabidopsis. Biol Chem. 2023 Sep 29;404(11-12):1137-1149.doi: 10.1515/hsz-2023-0195.
    Chung et al. (2023). An RNA thermometer in the chloroplast genome of Chlamydomonas facilitates temperature-controlled gene expression. Nucleic Acids Res. 2023 Nov 10;51(20):11386-11400. doi: 10.1093/nar/gkad816.
    Shi et al. (2023). Protocol to identify protein-protein interaction networks in Solanum tuberosum using transient TurboID-based proximity labeling. STAR Protoc. 2023 Sep 20;4(4):102577.doi: 10.1016/j.xpro.2023.102577.
    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. Miloro et al. (2024). Barley AGO4 proteins show overlapping functionality with distinct small RNA-binding properties in heterologous complementation. Plant Cell Rep. 2024 Mar 13;43(4):96. doi: 10.1007/s00299-024-03177-z.  
    Liu et al. (2023). RBPome identification in egg-cell like callus of Arabidopsis. Biol Chem. 2023 Sep 29;404(11-12):1137-1149.doi: 10.1515/hsz-2023-0195.
    Chung et al. (2023). An RNA thermometer in the chloroplast genome of Chlamydomonas facilitates temperature-controlled gene expression. Nucleic Acids Res. 2023 Nov 10;51(20):11386-11400. doi: 10.1093/nar/gkad816.
    Shi et al. (2023). Protocol to identify protein-protein interaction networks in Solanum tuberosum using transient TurboID-based proximity labeling. STAR Protoc. 2023 Sep 20;4(4):102577.doi: 10.1016/j.xpro.2023.102577.
    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.
  • Protocols
  • Agrisera Western Blot protocol and video tutorials
  • Reviews:
  • Reviews
    Below, you can grade the product on a scale from 0 to 5.
    Please also provide information about species, application, dilution and obtained result for the reviewed antibody.
    Your name will be displayed as the sender.
    Number of reviews: (12)
    Shelby Hotz | 2023-03-21
    0 | 0
    For my lab's study, we used this antibody for western blots to detect the abundance of Hsp70 in heat shocked and non-heat shocked bull kelp (Nereocystis leutkeana). We used a 1:5000 for the initial dilution and then a 1:2500 dilution after adding glycerol. Both were done in a 5% Nonfat Dried Milk in 1x Tris Buffered Saline solution. The results were positive, with the protein bands showing up clearly and at the correct weight according to the positive control we used.I am happy to report the antibody has been working effectively and as intended in our molecular research. The instructions that come with the product are very comprehensive and relatively easy to follow, and doing so has allowed us to effectively use the materials as expected and needed. We also appreciate the promptness of the delivery and arrival of the materials, and having the antibody in a lyophilized form is very much appreciated.Thank you for your service.
    Vasundara Sundar | 2022-12-28
    0 | 0
    We are very satisfied with the goat anti-rabbit antibodies, it worked well with all of the primary antibodies we used. 1:10000 dilution worked well for us.
    Annika Brünje | 2021-01-19
    0 | 0
    This antibody gives very good results in my Western blot analyses and can be also used in combination with primary antibodies that are a bit more tricky in their application. Usually, I use it in a dilution of 1:10000 to detect proteins in Arabidopsis leaf extracts.
    Soo Yeon Ko | 2020-05-28
    0 | 0
    I was satisfied with goat anti-rabbit IgG antibodies on protein extracts from Oryza Sativa in dilution : 1: 25 000
    ?udmila Holubová | 2019-10-04
    0 | 0
    We are happy with this antibody, works well with all primary antibodies we used. Our dilution was 1:12500.
    Ella Nukarinen | 2018-05-23
    0 | 0
    We are very happy with goat anti-rabbit IgG HRP antibody. We use it for Western blots in dilution 1:10 000-1:20 000 for detecting endogenous proteins from Arabidopsis protein extracts.
    Beata Kmiec | 2017-02-13
    0 | 0
    We're very happy with the antibodies. Our regular application is in western blots with Arabidopsis thaliana total protein extract and with isolated mitochondria and chloroplasts, but we've used it with success in WB with yeast and human samples. We use it in dilution 1:20000.
    Malgorzata Kwasniak-Owczarek | 2016-11-02
    0 | 0
    We are very satisfied with goat anti-rabbit IgG antibodies on protein extracts from Arabidopsis thaliana in dilution: 1: 25 000.
    Mirka Herbstova | 2016-04-22
    0 | 0
    I used Goat-anti rabbit IgG HRP conjugated secondary antibody (AS 09 602) at a dilution 1:50 000 for alga species Phaeodactylum and Trachydiscus without cross-reactions.
    Jakub Dolata, Adam Mickiewicz University, Poznan, Poland | 2014-03-12
    0 | 0
    The goat anti-rabbit IgG-HRP from Agrisera are very nice working antibodies with no unspecific signals in different dillutions (1:50000, 1:100000). We are using them in case of plant (Arabidopsis, Solanum) and human cells extracts as well.
    Jakub DOlata | 2014-03-12
    0 | 0
    The goat anti-rabbit IgG-HRP from Agrisera are very nice working antibodies with no unspecific signals in different dillutions (1:50000
    Gerhard Obermeyer, Univ Salzburg, Austria | 2011-11-11
    0 | 0
    We were looking for a 2nd anti-rabbit antibody which does not cross-react with proteins from a crude lily pollen lysate. The goat anti-rabbit IgG-HRP from Agrisera was the only one which showed no unspecific signals and could be used in a 1:40,000 dilution. All other antibodies from 5 different sources cross-reacted with a high number of proteins in the pollen lysate.

Accessories

img missing
AS16 ECL-N |  low pico to mid femtogram detection

This product can be purchased in 3 different volumes:

AS16 ECL-N-10, 10 ml. Trial size limited to one per customer

AS16 ECL-N-100, 100 ml

AS16 ECL-N-1L, 1L

Choose the appropriate volume in the drop down menu to the right

From 13 €
img missing
AS16 ECL-S | extreme low femtogram detection

This product can be purchased in 3 different volumes:

AS16 ECL-S-10, 10 ml. Trial size limited to one per customer

AS16 ECL-S-100, 100 ml

AS16 ECL-S-1L, 1L

Choose the appropriate volume in the drop down menu to the right

From 17 €
Close