BSA and streptavidin were from Sigma-Aldrich. PBS was from Gibco. TT was supplied by the Statens Serum Institut. All recombinant influenza HA subtypes were from Sino Biological: H1N1 HA (A/California/04/2009; H1), H2N2 HA (A/Japan/305/1957; H2), H5N1 HA (A/Vietnam/1194/2004; H5), H3N2 HA (A/Perth/16/2009; H3) and H7N9 HA (A/Shanghai/1/2013; H7). H1N1 HA (A/New Caledonia/20/1999; H1 wt), the HA delta stem mutants H1 I45R/T49R and H1 45glyc, and the human monoclonal antibodies CR6261 and CH65 were provided by D. Lingwood. HIV-1 92BR-gp120 was provided by D.R. Burton (Scripps Institute, San Diego, CA). Human CpG ODN 2006 was from InvivoGen. IL-15 and IL-6 were from PeproTech. Antibodies used were as follows: goat anti–human IgG (FcΥ specific; Jackson ImmunoResearch Laboratories, Inc.); anti–human CD27-APC, anti–human CD38-FITC, CD138-PE, CD19-Percp Cy5.5, CD20-APC, CD80-PE-Cy5, HLA-DR-PE-Cy5, and anti–human κ-Ig biotin (eBioscience); anti-human CD86-Bv510, CCR10-PE, CD62L, CXCR4-PE-Cy5, CXCR5-APC, and anti–human λ-Ig biotin (BioLegend); and anti–human CD43-PE and anti–human IgD (Miltenyi Biotec).
H5n1 ha
Manufactured by Sino Biological
Sourced in Japan
The H5N1 HA is a recombinant protein that corresponds to the hemagglutinin (HA) surface glycoprotein of the H5N1 influenza virus. It can be used for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
H3n2 ha, by Sino Biological (2 mentions)
H1n1 ha, by Sino Biological (2 mentions)
Cd138 pe, by Thermo Fisher Scientific (1 mentions)
Cd80 pe cy5, by Thermo Fisher Scientific (1 mentions)
Cd19 percp cy5, by Thermo Fisher Scientific (1 mentions)
Pe ccr10, by BioLegend (1 mentions)
Cxcr5 apc, by BioLegend (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Hla dr pe cy5, by Thermo Fisher Scientific (1 mentions)
Cd20 apc, by Thermo Fisher Scientific (1 mentions)
Cd62l, by BioLegend (1 mentions)
Hrp conjugated goat anti mouse igg h l, by Southern Biotech (1 mentions)
Igg2a, by Southern Biotech (1 mentions)
2 protocols using h5n1 ha
1
Binding Interactions of Influenza Hemagglutinin Subtypes
2
ELISA for Flu Antigen Antibodies
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The titers of serum IgG, IgG1, and IgG2a were measured
using an enzyme-linked
immunosorbent assay (ELISA). Briefly, 1 μg/mL of each flu antigen,
including H1N1 HrHA (derived from A/Puerto Rico/8/1934), H1N1 HA (A/California/04/2009),
H3N2 HA (A/Aichi/2/1968) (SinoBiological), H5N1 HA (A/Vietnam/1194/2004)
(SinoBiological), and tZE-4M2e, in PBS was coated onto Maxisorp 96
well immune assay plates during overnight incubation at 25 °C.
For the evaluation of off-target immune responses, Maxisorp 96-well
plates were coated with 1 μg/mL of tGCN4, tZR, or ZE-mCherry.
The next day, each well was washed three times with PBS containing
0.1% Tween-20 and blocked with 1% BSA-supplemented PBS for 2 h at
25 °C. Each well was incubated with serially diluted sera for
1 h at 25 °C and washed three times. Then, a 1:5000 dilution
of HRP-conjugated goat antimouse IgG (H+L) (SouthernBiotech), IgG1
(SouthernBiotech), or IgG2a (SouthernBiotech) secondary antibodies
was added to each well, incubated for 1 h at 25 °C, and washed
three times. Each well was developed with 1-Step Ultra TMB substrate
solution for 15 min and stopped with 2 N H2SO4 solution. Absorbance values at 450 nm were measured on a BioTek
Synergy H4 Micro plate reader with the correction wavelength set to
540 nm.
using an enzyme-linked
immunosorbent assay (ELISA). Briefly, 1 μg/mL of each flu antigen,
including H1N1 HrHA (derived from A/Puerto Rico/8/1934), H1N1 HA (A/California/04/2009),
H3N2 HA (A/Aichi/2/1968) (SinoBiological), H5N1 HA (A/Vietnam/1194/2004)
(SinoBiological), and tZE-4M2e, in PBS was coated onto Maxisorp 96
well immune assay plates during overnight incubation at 25 °C.
For the evaluation of off-target immune responses, Maxisorp 96-well
plates were coated with 1 μg/mL of tGCN4, tZR, or ZE-mCherry.
The next day, each well was washed three times with PBS containing
0.1% Tween-20 and blocked with 1% BSA-supplemented PBS for 2 h at
25 °C. Each well was incubated with serially diluted sera for
1 h at 25 °C and washed three times. Then, a 1:5000 dilution
of HRP-conjugated goat antimouse IgG (H+L) (SouthernBiotech), IgG1
(SouthernBiotech), or IgG2a (SouthernBiotech) secondary antibodies
was added to each well, incubated for 1 h at 25 °C, and washed
three times. Each well was developed with 1-Step Ultra TMB substrate
solution for 15 min and stopped with 2 N H2SO4 solution. Absorbance values at 450 nm were measured on a BioTek
Synergy H4 Micro plate reader with the correction wavelength set to
540 nm.
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