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Goat anti-Chicken IgY (H&L), HRP conjugated

AS09 603 | Clonality: Polyclonal  |  Host: Goat  | Reactivity: Hen IgY (H&L)

10 % until end of 2023. Use discount code: Conj10

Goat anti-Chicken IgY (H&L), HRP conjugated in the group Secondary Antibodies / Anti-Chicken  / HRP (horse radish peroxidase) at Agrisera AB (Antibodies for research) (AS09 603)
Goat anti-Chicken IgY (H&L), HRP conjugated



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Product Information

Immunogen Purified chicken IgY, whole molecule
Host Goat
Clonality Polyclonal
Purity Immunogen affinity purified goat 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. 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 : 10 000 -1 : 150 000 (ELISA), 1 : 500 -1 : 5000 (IHC), 1 : 20 000 and 1 : 10 000 (WB)

Reactivity

Confirmed reactivity Chicken IgY heavy and light chains (H&L).
Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples

Application example

western blot using Agrisera hen anti-PsbA antibody and Agrisera goat anti-hen secondary HRP conjugated

5 µg of total extract from (1) Hordeum vulgare total 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-PsbA antibody (AS01 016) 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 (anti-hen IgG horse radish peroxidase conjugated, 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 chemiluminescen 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. 

 


western blot detection of plant BiP using hen antibody

5 µg of total protein from  A.thaliana (1), H. vulgare (2) Z.mays (3) S. oleracea (4), extracted with Agrisera PEB extraction buffer (AS08 300) were separated on  4-12% SDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in  for 1h at room temperature with agitation. Blots were incubated in the primary antibody 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 (anti-hen IgY horse radish peroxidase conjugated, from  Agrisera AS09 603) diluted to 1:50 000  for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL  detection reagent according to the manufacturers instructions.  Exposure time was 5 seconds.

Additional information

Additional information

Concentration: 1.0 mg/ml

This HRP-conjugate is supplied in 10 mM Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 1 % (w/v) BSA, Protease/IgG free 0.1 % (v/v) of Kathon CG is used as preservative.

Antibody binds to:
heavy chains on chicken IgY
light chains on all chicken immunoglobulins

No reactivity is observed to non-immunoglobulin chicken serum proteins based in immunoelectrophoresis.

Chicken immunoglobulin is often called hen or chicken IgG, however it is derived from egg yolk, therefore IgY.

Related products

Background

Background

Goat anti-chicken IgY (H&L) is a secondary antibody conjugated to HRP (horse radish peroxidase) which binds to chicken IgY in immunological assays.

Product citations

Selected references Bindari et al. (2020). Methods to prevent PCR amplification of DNA from non-viable virus were not successful for infectious laryngotracheitis virus. PLoS One. 2020 May 22;15(5):e0232571. doi: 10.1371/journal.pone.0232571. PMID: 32442180; PMCID: PMC7244108.
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.
Heard et al. (2015). Identification of Regulatory and Cargo Proteins of Endosomal and Secretory Pathways in Arabidopsis thaliana by Proteomic Dissection. Mol Cell Proteomics. 2015 Jul;14(7):1796-813. doi: 10.1074/mcp.M115.050286. Epub 2015 Apr 21.
Huang et al. (2015). Effects of exogenous salicylic acid on the physiological characteristics of Dendrobium officinale under chilling stress. Plant Growth Regulation pp 1-10.
immunogen: Purified chicken IgY, whole molecule
Host: Goat
Clonality: Polyclonal
Purity: Immunogen affinity purified goat 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. 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 : 10 000 -1 : 150 000 (ELISA), 1 : 500 -1 : 5000 (IHC), 1 : 20 000 and 1 : 10 000 (WB)
Confirmed reactivity: Chicken IgY heavy and light chains (H&L).
not reactive in: No confirmed exceptions from predicted reactivity are currently known
Picture (footer):

Application example

western blot using Agrisera hen anti-PsbA antibody and Agrisera goat anti-hen secondary HRP conjugated

5 µg of total extract from (1) Hordeum vulgare total 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-PsbA antibody (AS01 016) 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 (anti-hen IgG horse radish peroxidase conjugated, 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 chemiluminescen 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. 

 


western blot detection of plant BiP using hen antibody

5 µg of total protein from  A.thaliana (1), H. vulgare (2) Z.mays (3) S. oleracea (4), extracted with Agrisera PEB extraction buffer (AS08 300) were separated on  4-12% SDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in  for 1h at room temperature with agitation. Blots were incubated in the primary antibody 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 (anti-hen IgY horse radish peroxidase conjugated, from  Agrisera AS09 603) diluted to 1:50 000  for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with ECL  detection reagent according to the manufacturers instructions.  Exposure time was 5 seconds.

additional information:

Concentration: 1.0 mg/ml

This HRP-conjugate is supplied in 10 mM Sodium Phosphate, 0.15 M Sodium Chloride, pH 7.2, 1 % (w/v) BSA, Protease/IgG free 0.1 % (v/v) of Kathon CG is used as preservative.

additional information (application):

Antibody binds to:
heavy chains on chicken IgY
light chains on all chicken immunoglobulins

No reactivity is observed to non-immunoglobulin chicken serum proteins based in immunoelectrophoresis.

Chicken immunoglobulin is often called hen or chicken IgG, however it is derived from egg yolk, therefore IgY.

background:

Goat anti-chicken IgY (H&L) is a secondary antibody conjugated to HRP (horse radish peroxidase) which binds to chicken IgY in immunological assays.

All references: Bindari et al. (2020). Methods to prevent PCR amplification of DNA from non-viable virus were not successful for infectious laryngotracheitis virus. PLoS One. 2020 May 22;15(5):e0232571. doi: 10.1371/journal.pone.0232571. PMID: 32442180; PMCID: PMC7244108.
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.
Heard et al. (2015). Identification of Regulatory and Cargo Proteins of Endosomal and Secretory Pathways in Arabidopsis thaliana by Proteomic Dissection. Mol Cell Proteomics. 2015 Jul;14(7):1796-813. doi: 10.1074/mcp.M115.050286. Epub 2015 Apr 21.
Huang et al. (2015). Effects of exogenous salicylic acid on the physiological characteristics of Dendrobium officinale under chilling stress. Plant Growth Regulation pp 1-10.

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