Microcrystalline wax (Ibercer 2616, melting point of 81 °C) was kindly provided by Iberceras Specialities S.L.U. (Madrid, Spain). We purchased 100 µm-thick polyester transparency A4 sheets from APLI S.L. (Alaquàs, Spain). Pressure-sensitive-adhesive films (PSA) (ARcare® 8939, 120 µm total thickness) were obtained from Adhesives Research Inc. (Glen Rock, PA, USA). Poly(methyl methacrylate) (PMMA) was acquired from Servicio Estación (Barcelona, Spain). Unbacked 120 µm-thick nitrocellulose was obtained from GE Healthcare Life Sciences (Piscataway, NJ, USA). Polydimethylsiloxane (PDMS Sylgard 184 kit, which includes the Sylgard 184 silicone elastomer base and the Sylgard 184 silicone elastomer-curing agent) was purchased from Sigma Aldrich (St. Louis, MO, USA). For the immunoassay, the capture (anti-human) and detector antibodies (anti-human-HRP), along with the analyte (TNFα), were obtained from Sinobiological Inc. (Beijing, China). The buffer reagents (Phosphate Buffered Saline (PBS), Tween 20, Bovine Serum Albumin (BSA)), the dyes (Tartrazine, Erioglaucine and Allura red), the streptavidine-HRP, as well as the substrate (3,3′,5,5′-Tetramethylbenzidine (TMB)) were obtained from Sigma Aldrich (USA). Sodium Hydrogen Carbonate (NaHCO3) was purchased from Panreac Química (Barcelona, Spain).
Antihuman hrp
Manufactured by Sino Biological
Sourced in China
Antihuman-HRP is a horseradish peroxidase (HRP)-conjugated secondary antibody that binds to human antibodies. It is commonly used in various immunoassays and detection techniques to amplify and visualize the signal from human antibodies.
Automatically generated - may contain errors
Lab products found in correlation
Erioglaucine, by Merck Group (1 mentions)
Tnf α, by Sino Biological (1 mentions)
Streptavidine hrp, by Merck Group (1 mentions)
Allura red, by Merck Group (1 mentions)
Tartrazine, by Merck Group (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Maxisorp, by Thermo Fisher Scientific (1 mentions)
Tmb substrate, by Thermo Fisher Scientific (1 mentions)
3 protocols using antihuman hrp
1
Fabrication of Microcrystalline Wax Immunoassay
2
SARS-CoV-2 Spike-Heparin Binding Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Nunc maxisorp flat bottom 96 well
plates were coated with streptavidin (200 nM; 50 μL) overnight
at 4 °C. The plates were blocked with 2% BSA for 1 h and biotinylated
heparin (800 nM; 50 μL) was added to the plates for 1 h. Unbound
GAGs were thoroughly washed three times with 200 μL of 1×PBST
(0.05% Tween 20). Variant spike proteins along with wild-type spike
protein (100 nM; 50 μL) diluted in the kinetic buffer (10 mM
HEPES, 10 mM NaCl, 0.05% Tween 20, pH 7.4) were added to the plate
and incubated for 1 h. Unbound spike protein was washed three times
with 1×PBST and NTD Abs (2 μg/mL; 50 μL) diluted
in the kinetic buffer were added for sandwich-type binding. Unbound
NTD Abs were washed three times with 1×PBST and incubated with
50 μL each of 0.1 μg/mL antihuman-HRP (Sino Biological,
10702-T16-H) for 30 min at room temperature. The wells were washed
thoroughly 5 times with 200 μL of 1×PBST. Finally, 100
μL of TMB substrate (Thermo Fisher Scientific, 34028 Thermo
Fisher Scientific, 34028) was added to each well to develop color.
The reaction was stopped by adding 50 μL of stop solution (Thermo
Fisher Scientific, N600), and absorbance was measured at 450 nm.
plates were coated with streptavidin (200 nM; 50 μL) overnight
at 4 °C. The plates were blocked with 2% BSA for 1 h and biotinylated
heparin (800 nM; 50 μL) was added to the plates for 1 h. Unbound
GAGs were thoroughly washed three times with 200 μL of 1×PBST
(0.05% Tween 20). Variant spike proteins along with wild-type spike
protein (100 nM; 50 μL) diluted in the kinetic buffer (10 mM
HEPES, 10 mM NaCl, 0.05% Tween 20, pH 7.4) were added to the plate
and incubated for 1 h. Unbound spike protein was washed three times
with 1×PBST and NTD Abs (2 μg/mL; 50 μL) diluted
in the kinetic buffer were added for sandwich-type binding. Unbound
NTD Abs were washed three times with 1×PBST and incubated with
50 μL each of 0.1 μg/mL antihuman-HRP (Sino Biological,
10702-T16-H) for 30 min at room temperature. The wells were washed
thoroughly 5 times with 200 μL of 1×PBST. Finally, 100
μL of TMB substrate (Thermo Fisher Scientific, 34028 Thermo
Fisher Scientific, 34028) was added to each well to develop color.
The reaction was stopped by adding 50 μL of stop solution (Thermo
Fisher Scientific, N600), and absorbance was measured at 450 nm.
3
SARS-CoV-2 Spike Protein Binding Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Nunc maxisorp flat bottom 96 well plates were coated
with streptavidin (200 nM; 50 μL) overnight at 4 °C. The
plates were blocked with 2% BSA for 1 h and biotinylated GAGs (800
nM; 50 μL) were added to the plates for 1 h. Unbound GAGs were
thoroughly washed with 200 μL of 1xPBST (0.05% Tween 20) for
three times. Spike proteins (100 nM; 50 μL) diluted in the kinetic
buffer (10 mM HEPES, 10 mM NaCl, 0.05% Tween 20, pH 7.4) were added
to the plate and incubated for 1 h. Unbound spike protein were washed
three times with 1xPBST and NTD Abs (2 μg/mL; 50 μL) diluted
in the kinetic buffer were added for sandwich-type binding. Unbound
NTD Abs were washed three times with 1×PBST and incubated with
50 μL each of 0.1 μg/mL antihuman-HRP (Sino Biological,
10702-T16-H) for 30 min at room temperature. The wells were washed
thoroughly 5 times with 200 μL of 1×PBST. Finally, 100
μL of TMB substrate (Thermo Fisher Scientific, 34028) was added
to each well to develop color. The reaction was stopped by adding
50 μL of stop solution (Thermo Fisher Scientific, N600) and
absorbance was measured at 450 nm.
with streptavidin (200 nM; 50 μL) overnight at 4 °C. The
plates were blocked with 2% BSA for 1 h and biotinylated GAGs (800
nM; 50 μL) were added to the plates for 1 h. Unbound GAGs were
thoroughly washed with 200 μL of 1xPBST (0.05% Tween 20) for
three times. Spike proteins (100 nM; 50 μL) diluted in the kinetic
buffer (10 mM HEPES, 10 mM NaCl, 0.05% Tween 20, pH 7.4) were added
to the plate and incubated for 1 h. Unbound spike protein were washed
three times with 1xPBST and NTD Abs (2 μg/mL; 50 μL) diluted
in the kinetic buffer were added for sandwich-type binding. Unbound
NTD Abs were washed three times with 1×PBST and incubated with
50 μL each of 0.1 μg/mL antihuman-HRP (Sino Biological,
10702-T16-H) for 30 min at room temperature. The wells were washed
thoroughly 5 times with 200 μL of 1×PBST. Finally, 100
μL of TMB substrate (Thermo Fisher Scientific, 34028) was added
to each well to develop color. The reaction was stopped by adding
50 μL of stop solution (Thermo Fisher Scientific, N600) and
absorbance was measured at 450 nm.
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