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Kpl stop solution

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The KPL stop solution is a laboratory product designed to halt enzymatic reactions. It is a commonly used reagent in various analytical and experimental protocols.

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Lab products found in correlation

Maxisorp plate, by Thermo Fisher Scientific (15 mentions) Kpl tmb 2 component peroxidase substrate kit, by LGC (14 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (12 mentions) Goat anti hamster igg fc, by Abcam (5 mentions) Blocker casein, by Thermo Fisher Scientific (1 mentions) Pbs blocking buffer, by Thermo Fisher Scientific (1 mentions)

15 protocols using kpl stop solution

1

ELISA Assay for SARS-CoV-2 Antibodies

Cited 1 time
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ELISAs were
performed as previously described.22 (link) In
brief, Maxisorp plates (Nunc) were coated with 50 ng of spike protein
(Sinobiological SARS-CoV-2 (2019-nCoV) spike S1 + S2 ECD-HIS recombinant
protein (catalog #40589-V08B1)) per well. Plates were incubated overnight
at 4 °C. Plates were blocked with Blocker casein in phosphate
buffered saline (PBS) (ThermoFisher) for 1 h at room temperature (RT).
Hamster serum collected on day 14 post exposure was diluted 1:400
in Blocker casein in PBS and incubated for 1 h at RT. Serum from a
subset of positive qRT-PCR animals was titrated via a 2× serial
dilution to obtain antibody titers of positive animals. Secondary
goat antihamster IgG Fc (horseradish peroxidase-conjugated, Abcam)
spike-specific antibodies were used for detection and visualized with
a KPL TMB 2-component peroxidase substrate kit (SeraCare, 5120-0047).
The reaction was stopped with KPL stop solution (SeraCare), and the
plates were read at 450 nm. The threshold for positivity was calculated
as the average plus 3 × the standard deviation of prebleed serum
from three animals as a negative control.
Fischer R.J., Port J.R., Holbrook M.G., Yinda K.C., Creusen M., ter Stege J., de Samber M, & Munster V.J. (2022). UV-C Light Completely Blocks Aerosol Transmission of Highly Contagious SARS-CoV-2 Variants WA1 and Delta in Hamsters. Environmental Science & Technology, 56(17), 12424-12430.
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2

SARS-CoV-2 Spike Protein Antibody Assay

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Serum samples were inactivated with γ-irradiation (2 mRad) and analyzed as previously described [63 ]. In brief, maxisorp plates (Nunc) were coated with 50 ng spike protein (generated in-house) per well and incubated overnight at 4°C. After blocking with casein in phosphate buffered saline (PBS) (ThermoFisher) for 1 h at room temperature (RT), serially diluted 2-fold serum samples (duplicate, in blocking buffer) were incubated for 1 h at RT. Spike-specific antibodies were detected with goat anti-hamster IgG Fc (horseradish peroxidase (HRP)-conjugated, Abcam) for 1 h at RT and visualized with KPL TMB 2-component peroxidase substrate kit (SeraCare, 5120–0047). The reaction was stopped with KPL stop solution (Seracare) and read at 450 nm. Plates were washed 3 to 5 × with PBS-T (0.1% Tween) for each wash. The threshold for positivity was calculated as the average plus 3 × the standard deviation of negative control hamster sera.
Port J.R., Yinda C.K., Owusu I.O., Holbrook M., Fischer R., Bushmaker T., Avanzato V.A., Schulz J.E., van Doremalen N., Clancy C.S, & Munster V.J. (2020). SARS-CoV-2 disease severity and transmission efficiency is increased for airborne but not fomite exposure in Syrian hamsters.. bioRxiv.
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3

SARS-CoV-2 Spike Protein Antibody Assay

Cited 1 time
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Serum samples were inactivated with γ-irradiation (2 mRad) and analyzed as previously described [28 (link)]. In brief, maxisorp plates (Nunc) were coated with 50 ng spike protein (generated in-house) per well and incubated overnight at 4 °C. After blocking with casein in phosphate buffered saline (PBS) (ThermoFisher) for 1 h at room temperature (RT), serially diluted 2-fold serum samples (duplicate, in blocking buffer) were incubated for 1 h at RT. Spike-specific antibodies were detected with goat anti-hamster IgG Fc (horseradish peroxidase (HRP)-conjugated, Abcam) for 1 hr at RT and visualized with KPL TMB 2-component peroxidase substrate kit (SeraCare, 5120-0047, Milford, MA, USA). The reaction was stopped with KPL stop solution (Seracare) and read at 450 nm. Plates were washed 3 to 5× with PBS-T (0.1% Tween) for each wash. The threshold for positivity was calculated as the average plus 3× the standard deviation of negative control hamster sera.
Port J.R., Adney D.R., Schwarz B., Schulz J.E., Sturdevant D.E., Smith B.J., Avanzato V.A., Holbrook M.G., Purushotham J.N., Stromberg K.A., Leighton I., Bosio C.M., Shaia C, & Munster V.J. (2021). High-Fat High-Sugar Diet-Induced Changes in the Lipid Metabolism Are Associated with Mildly Increased COVID-19 Severity and Delayed Recovery in the Syrian Hamster. Viruses, 13(12), 2506.
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4

SARS-CoV-2 Spike Protein Antibody ELISA

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Maxisorp plates (Nunc) were coated with 50 ng spike protein per well and incubated overnight at 4°C. After blocking with casein in phosphate buffered saline (PBS) (ThermoFisher) for 1 h at room temperature (RT), serially diluted 2-fold serum samples (duplicate, in casein) were incubated for 1 h at RT. Spike-specific antibodies were detected with goat anti-mouse IgM or IgG Fc (horseradish peroxidase (HRP)-conjugated, Abcam) for 1 h at RT and visualized with KPL TMB 2-component peroxidase substrate kit (SeraCare, 5120–0047). The reaction was stopped with KPL stop solution (Seracare) and read at 450 nm. Plates were washed 3x with PBS-T (0.1% Tween) in between steps. The threshold for positivity was calculated as the average plus 3x the standard deviation of negative control mouse sera.
Yinda C.K., Port J.R., Bushmaker T., Offei Owusu I., Purushotham J.N., Avanzato V.A., Fischer R.J., Schulz J.E., Holbrook M.G., Hebner M.J., Rosenke R., Thomas T., Marzi A., Best S.M., de Wit E., Shaia C., van Doremalen N, & Munster V.J. (2021). K18-hACE2 mice develop respiratory disease resembling severe COVID-19. PLoS Pathogens, 17(1), e1009195.
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5

SARS-CoV-2 Spike Protein Antibody Assay

Cited 1 time
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Serum samples were analyzed as previously described [57 (link)]. In brief, maxisorp plates (Nunc) were coated with 50 ng spike protein (generated in-house) per well. Plates were incubated overnight at 4°C. Plates were blocked with casein in phosphate buffered saline (PBS) (ThermoFisher) for 1 hour at room temperature. Serum was diluted 2-fold in blocking buffer and samples (duplicate) were incubated for 1 hour at room temperature. Secondary goat anti-hamster IgG Fc (horseradish peroxidase (HRP)-conjugated, Abcam) spike-specific antibodies were used for detection and visualized with KPL TMB 2-component peroxidase substrate kit (SeraCare, 5120–0047). The reaction was stopped with KPL stop solution (Seracare) and plates were read at 450 nm. The threshold for positivity was calculated as the average plus 3 × the standard deviation of negative control hamster sera.
Port J.R., Yinda C.K., Riopelle J.C., Weishampel Z.A., Saturday T.A., Avanzato V.A., Schulz J.E., Holbrook M.G., Barbian K., Perry-Gottschalk R., Haddock E., Martens C., Shaia C.I., Lambe T., Gilbert S.C., van Doremalen N, & Munster V.J. (2022). Infection- or vaccine mediated immunity reduces SARS-CoV-2 transmission, but increases competitiveness of Omicron in hamsters. bioRxiv.
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6

SARS-CoV-2 Spike Protein ELISA Assay

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Purified SARS-CoV-2 full length spike or RBD protein was diluted to 1 μg/mL in PBS. Maxisorp plates (Nunc) were coated with 100 μL per well (100 ng protein per well) and incubated overnight at 4°C. Plates were washed 3x with PBST (0.1% Tween) and blocked with 100 μL casein in PBS blocking buffer (ThermoFisher) for 1 hour at room temp. Plates were again washed 3x with PBST (0.1% Tween), and 100 μL of serum samples, serially diluted 2 fold in casein in PBS blocking buffer, in duplicate, was added to the wells and incubated at room temperature for 1 hour. Plates were washed 4x with PBST (0.1% Tween), and 100 μL secondary antibody, rabbit anti-human IgG Fc HRP (Novus Biologicals, NBP1-73529) diluted 1:4000 in casein in PBS blocking buffer, was added to the wells and incubated for 1 hour at room temperature. The wells were washed 5x with PBST (0.1% Tween) and developed with the KPL TMP 2-component peroxidase substrate kit (Seracare, 5120-0047). The reaction was stopped with KPL stop solution (Seracare, 5150-0020) and read at 450 nm. The threshold for positivity was calculated as the average plus 3 times the standard deviation of negative control sera. Reported titers are the reciprocal value of the highest dilution at which signal was observed above the calculated threshold.
Avanzato V.A., Matson M.J., Seifert S.N., Pryce R., Williamson B.N., Anzick S.L., Barbian K., Judson S.D., Fischer E.R., Martens C., Bowden T.A., de Wit E., Riedo F.X, & Munster V.J. (2020). Case Study: Prolonged Infectious SARS-CoV-2 Shedding from an Asymptomatic Immunocompromised Individual with Cancer. Cell, 183(7), 1901-1912.e9.
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7

SARS-CoV-2 Spike Protein Antibody Assay

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Serum samples were analyzed as previously described55 (link). In brief, maxisorp plates (Nunc) were coated with 50 ng spike protein (generated in-house) per well. Plates were incubated overnight at 4°C. Plates were blocked with casein in phosphate buffered saline (PBS) (ThermoFisher) for 1 hours at room temperature (RT). Serum was diluted 2-fold in blocking buffer and samples (duplicate) were incubated for 1 hours at RT. Secondary goat anti-hamster IgG Fc (horseradish peroxidase (HRP)-conjugated, Abcam) spike-specific antibodies were used for detection and visualized with KPL TMB 2-component peroxidase substrate kit (SeraCare, 5120–0047). The reaction was stopped with KPL stop solution (Seracare) and plates were read at 450 nm. The threshold for positivity was calculated as the average plus 3 × the standard deviation of negative control hamster sera.
Port J.R., Yinda C.K., Avanzato V.A., Schulz J.E., Holbrook M.G., van Doremalen N., Shaia C., Fischer R.J, & Munster V.J. (2021). Increased aerosol transmission for B.1.1.7 (alpha variant) over lineage A variant of SARS-CoV-2. bioRxiv.
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8

SARS-CoV-2 Spike Protein IgG ELISA

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Serum samples were analyzed as previously described (Yinda et al., 2021 (link)). In brief, maxisorp plates (Nunc) were coated with 50 ng spike protein (generated in-house, purified recombinant) per well. Plates were incubated overnight at 4 °C. Plates were blocked with casein in phosphate buffered saline (PBS) (Thermo Fisher) for 1 hr at room temperature (RT). Serum was diluted twofold in blocking buffer and samples (duplicate) were incubated for 1 hr at RT. Secondary goat anti-hamster IgG Fc (horseradish peroxidase (HRP)-conjugated, Cat.No. 5220–0371 Lot. 10492253, Seracare) antibodies were used for detection and KPL TMB 2-component peroxidase substrate kit (SeraCare, Cat.No. 5120–0047) was used for visualization. The reaction was stopped with KPL stop solution (Seracare) and plates were read at 450 nm. The threshold for positivity was calculated as the average plus 3 x the standard deviation of negative control hamster sera.
Port J.R., Morris D.H., Riopelle J.C., Yinda C.K., Avanzato V.A., Holbrook M.G., Bushmaker T., Schulz J.E., Saturday T.A., Barbian K., Russell C.A., Perry-Gottschalk R., Shaia C., Martens C., Lloyd-Smith J.O., Fischer R.J, & Munster V.J. (2024). Host and viral determinants of airborne transmission of SARS-CoV-2 in the Syrian hamster. eLife, 12, RP87094.
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9

SARS-CoV-2 Spike Protein Antibody ELISA

Check if the same lab product or an alternative is used in the 5 most similar protocols
Maxisorp plates (Nunc) were coated with 50 ng spike protein per well and incubated overnight at 4°C. After blocking with casein in phosphate buffered saline (PBS) (ThermoFisher) for 1 h at room temperature (RT), serially diluted 2-fold serum samples (duplicate, in casein) were incubated for 1 h at RT. Spike-specific antibodies were detected with goat anti-mouse IgM or IgG Fc (horseradish peroxidase (HRP)-conjugated, Abcam) for 1 h at RT and visualized with KPL TMB 2-component peroxidase substrate kit (SeraCare, 5120–0047). The reaction was stopped with KPL stop solution (Seracare) and read at 450 nm. Plates were washed 3x with PBS-T (0.1% Tween) in between steps. The threshold for positivity was calculated as the average plus 3x the standard deviation of negative control mouse sera.
Yinda C.K., Port J.R., Bushmaker T., Owusu I.O., Avanzato V.A., Fischer R.J., Schulz J.E., Holbrook M.G., Hebner M.J., Rosenke R., Thomas T., Marzi A., Best S.M., de Wit E., Shaia C., van Doremalen N, & Munster V.J. (2020). K18-hACE2 mice develop respiratory disease resembling severe COVID-19. bioRxiv.
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10

SARS-CoV-2 Spike Protein Antibody Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Serum samples were inactivated with γ-irradiation (2 mRad) and analyzed as previously described (Yinda et al., 2020 ). In brief, maxisorp plates (Nunc) were coated with 50 ng spike protein (generated in-house) per well and incubated overnight at 4 °C. After blocking with casein in phosphate buffered saline (PBS) (ThermoFisher) for 1 h at room temperature (RT), serially diluted 2-fold serum samples (duplicate, in blocking buffer) were incubated for 1 h at RT. Spike-specific antibodies were detected with goat anti-hamster IgG Fc (horseradish peroxidase (HRP)-conjugated, Abcam) for 1 h at RT and visualized with KPL TMB 2-component peroxidase substrate kit (SeraCare, 5120–0047). The reaction was stopped with KPL stop solution (Seracare) and read at 450 nm. Plates were washed 3 to 5 × with PBS-T (0.1 % Tween) for each wash. The threshold for positivity was calculated as the average plus 3 × the standard deviation of negative control hamster sera.
Port J.R., Adney D.R., Schwarz B., Schulz J.E., Sturdevant D.E., Smith B.J., Avanzato V.A., Holbrook M.G., Purushotham J.N., Stromberg K.A., Leighton I., Bosio C.M., Shaia C, & Munster V.J. (2021). Western diet increases COVID-19 disease severity in the Syrian hamster. bioRxiv.
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