Immulon 2hb plate

Manufactured by Thermo Fisher Scientific

Sourced in United States

Immulon 2HB plates are 96-well polystyrene microplates designed for use in enzyme-linked immunosorbent assays (ELISA) and other immunoassay applications. The plates feature a high-binding surface that promotes the adsorption of protein antigens or antibodies. The 'HB' in the name stands for 'high-binding', indicating the enhanced protein-binding capacity of the plate surface.

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Lab products found in correlation

Epoch elisa reader, by Agilent Technologies (4 mentions) Anti human kappa ap, by Southern Biotech (4 mentions) Ap substrate, by Merck Group (4 mentions) Ifnαr2, by R&D Systems (4 mentions) Bovine serum albumin (bsa), by Southern Biotech (3 mentions) Hrp labeled goat anti mouse igg or igg2c fc antibody, by Thermo Fisher Scientific (3 mentions) Am9680, by Thermo Fisher Scientific (3 mentions) Bovine serum albumin (bsa), by Merck Group (3 mentions) Bovine serum albumin (bsa), by Thermo Fisher Scientific (2 mentions) Salmon sperm dna, by Thermo Fisher Scientific (2 mentions) Hep 2 slides, by MBL Life Science (2 mentions) Alexa flour 488 conjugated anti mouse igg, by Jackson ImmunoResearch (2 mentions) Ultra turrax vd125, by Avantor (2 mentions) Ultra tmb elisa substrate, by Thermo Fisher Scientific (2 mentions) Horseradish peroxidase hrp conjugated streptavidin, by Jackson ImmunoResearch (2 mentions) Goat anti human igg hrp, by Merck Group (2 mentions) Tmb substrate, by BD (2 mentions) Synergy htx multi mode microplate reader, by Agilent Technologies (2 mentions) Anti human igg isotype specific hrp, by Southern Biotech (2 mentions) Goat anti mouse hrp, by Jackson ImmunoResearch (1 mentions)

Close spelling variants of this product

Immulon 2HB (34 citations) Immulon 2HB 96-well plates (17 citations) Immulon 2HB microtiter plates (5 citations) Immulon 2HB ELISA plates (4 citations) Immulon 2HB 96-well microtiter plates (3 citations) IMMULON 2HB flat bottom plates (3 citations) Immulon 2HB flat-bottom 96-well plates (2 citations)

47 protocols using immulon 2hb plate

C1-INH Quantification via ELISA

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Immulon 2HB plates (Thermo Scientific, Waltham, Mass) were coated with 5 μg/mL polyclonal antibody to C1-INH. After blocking with 1% BSA in PBS, samples and standards were added and incubated at room temperature for 1 hour. Bound C1-INH was probed with alkaline phosphatase–conjugated mAb to C1-INH, followed by color development with 5-bromo-4-chloroindolyl phosphate/nitroblue tetrazolium (BCIP/NBT).

Joseph K., Tholanikunnel B.G., Wolf B., Bork K, & Kaplan A.P. (2015). Deficiency of plasminogen activator inhibitor 2 in plasma of patients with hereditary angioedema with normal C1 inhibitor levels. The Journal of allergy and clinical immunology, 137(6), 1822-1829.e1.

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Masked Fusion Abs Binding to IFNα2 Receptor

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Example 4

In this example, it is shown that a plurality of Masked Fusion Abs of the disclosure can bind to the IFNα2 receptor. Briefly, Immulon 2 HB plates (Thermofisher) were coated with 10 ug/mL IFNαR2 (R&D Systems) overnight at 4 deg. C and blocked with 2% BSA (Fisher) for a minimum of 2 hrs. at room temperature. Then, wells were washed 3× with PBS+0.05% Tween (Sigma). Indicated Ab concentrations were overlayed overnight at 4 deg. C. Wells were then washed 3× with PBS+0.05% Tween. Bound Abs were detected with anti-human Kappa-AP (Southern Biotech) diluted 1:3000 in PBS+1% BSA. Absorbance changes after addition of AP substrate (Sigma) were assayed at 410 nm using a Biotek EPOCH ELISA reader. The results show the masked 5T4, mesothelin, CD20, and CD138 Abs bind IFNαR2 with lower affinity compared to the unmasked 5T4, mesothelin, CD20, and CD138 Fusion Abs. (See, FIG. 4 and FIG. 5).

US11136353B2. Fusion protein composition(s) comprising masked type I interferons (IFNA and IFNB) for use in the treatment of cancer and methods thereof (2021-10-05). Qwixel Therapeutics LLC [US]. Inventors: David Stover [US], Sherie Morrison [US], Alex Vasuthasawat [US], Kham Trinh [US], George Ayoub [US].

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Masked Fusion Ab Binding to IFNαR2

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Example 20

In this example, it is shown that a Masked Fusion Ab (utilizing mask2) of the disclosure can bind to the IFNα2 receptor. Briefly, Immulon 2 HB plates (Thermofisher) were coated with 10 ug/mL IFNαR2 (R&D Systems) overnight at 4 deg. C and blocked with 2% BSA (Fisher) for a minimum of 2 hrs. at room temperature. Then, wells were washed 3× with PBS+0.05% Tween (Sigma). Indicated Ab concentrations were overlayed overnight at 4 deg. C. Wells were then washed 3× with PBS+0.05% Tween. Bound Abs were detected with anti-human Kappa-AP (Southern Biotech) diluted 1:3000 in PBS+1% BSA. Absorbance changes after addition of AP substrate (Sigma) were assayed at 410 nm using a Biotek EPOCH ELISA reader. The results show that both mask1 and mask2 are able to inhibit Fusion Ab binding to IFNαR2. (See, FIG. 31).

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Quantifying MNoV-Specific Antibody Levels

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Immulon 2HB plates (Thermo Fisher) were coated with UV inactivated MNoV virions or bacterially-produced MNoV NS1 (amino acids 28–114) in PBS. Plates were washed three times in ELISA wash buffer (0.15M NaCl, 0.05% Tween 20) and blocked with 3% bovine serum albumin. Serum was diluted in ELISA III buffer (0.15M NaCl, 0.001M EDTA, 0.05M Tris, 0.05% Tween 20, 0.1% BSA, pH 7.4) and added to plate for 1 hour at 37°C. Unbound serum antibody was removed by washing four times in ELISA wash buffer. Goat-anti-mouse-HRP (Jackson Immunoresearch) was diluted 1:1000 in ELISA III buffer and added to plate for 1 hour at 37°C. Plate was washed four times in ELISA wash buffer. HRP activity was detected by adding ELISA substrate buffer (0.1M sodium citrate, 1mM ABTS, 0.016% hydrogen peroxide) and stopping in 0.2N Phosphoric Acid. Absorbance was measured at 415nm.

Nice T.J., Osborne L.C., Tomov V.T., Artis D., Wherry E.J, & Virgin H.W. (2016). Type I Interferon Receptor Deficiency in Dendritic Cells Facilitates Systemic Murine Norovirus Persistence Despite Enhanced Adaptive Immunity. PLoS Pathogens, 12(6), e1005684.

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Peptide-Based ELISA for MPER and LACK

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MPER- and LACK-specific ELISAs were performed as before [35] (link). In brief, 96-Well Immulon 2HB Plates (Thermo Fisher Scientific) were coated overnight with streptavidin in PBS (50 µl/well) at 4 °C. Wells were washed, blocked, and coated with peptide/liposomes (palmitoylated MPER or LACK). Wells were washed again and incubated overnight at 4 °C with serially diluted sera. The following day, serum was removed, the plate was washed, incubated with HRP-conjugated secondary antibody, re-washed, the signal was developed with o-phenylenediamine (OPD) solution (see Supplement Table S5), and the absorbance was measured at 490 nm.

Elbahnasawy M.A., Donius L.R., Reinherz E.L, & Kim M. (2018). Co-delivery of a CD4 T cell helper epitope via covalent liposome attachment with a surface-arrayed B cell target antigen fosters higher affinity antibody responses. Vaccine, 36(41), 6191-6201.

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ELISA-based Malaria Serology Protocol

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Malaria serology was performed with an in-house screening ELISA for detection of specific Plasmodium spp. antibodies. P. falciparum antigen (strain NF54) was coated in 0.05 M sodium carbonate buffer, pH 9.6, to Immulon 2HB plates (, Thermo Scientific, Wohlen, Switzerland,). After washing, diluted sera were added to the plates and incubated for 15 min at 37 °C. After additional washing steps, horseradish peroxidase conjugated goat-anti-human-IgG (KPL, 474-1006, BioConcept Ltd, Allschwil, Switzerland) was added. Plates were incubated for 15 min at 37 °C, subsequently washed, and o-Phenylendiamine Dihydrochloride (OPD, Sigma, Buchs, Switzerland) was added. Reaction was stopped with 8 M H₂SO₄, and absorption was read with a Multiscan FC reader (ThermoScientific, Wohlen, Switzerland) at 492 nm. All sera giving positive or equivocal results were additionally tested with an in-house confirmatory Malaria IFA.

Mudji J., Blum A., Grize L., Wampfler R., Ruf M.T., Cnops L., Nickel B., Burri C, & Blum J. (2020). Gambiense Human African Trypanosomiasis Sequelae after Treatment: A Follow-Up Study 12 Years after Treatment. Tropical Medicine and Infectious Disease, 5(1), 10.

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Autoantibody Profiling by ELISA and Microarray

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For anti-dsDNA ELISA, plates were coated with 100 μg/mL salmon sperm DNA (Invitrogen; #AM9680) at 37 ^oC overnight and blocked in 2% BSA in PBS at room temperature for 2 h. For anti-cardiolipin and anti-phosphatidylserine ELISA, Immulon 2HB plates (Thermo) were coated with 75μg/mL of cardiolipin or with 30 μg/mL phosphatidylserine dissolved in 100% ethanol overnight. Sera were diluted 1:200 and incubated on coated plates at 25 ^oC for 2 h. Supernatants from sorted ABCs were used neat after 7 d culture with 1 μM Imiquimod. Plates were then incubated with HRP-labeled goat anti-mouse IgG or IgG2c Fc antibody for 1 h (eBioscience). OD₄₅₀ was measured on a microplate reader. Autoantibody activities against a panel of autoantigen specificities were measured using an Autoantigen Microarray platform developed by the University of Texas Southwestern Medical Center^{69 (link),70 (link)}.

Ricker E., Manni M., Flores-Castro D., Jenkins D., Gupta S., Rivera-Correa J., Meng W., Rosenfeld A.M., Pannellini T., Bachu M., Chinenov Y., Sculco P.K., Jessberger R., Prak E.T, & Pernis A.B. (2021). Altered function and differentiation of age-associated B cells contribute to the female bias in lupus mice. Nature Communications, 12, 4813.

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Autoantibody Detection Assays

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For anti-dsDNA ELISA, plates were coated with 100 Î¼g/ml salmon
sperm DNA (Invitrogen AM9680) at 37Â°C overnight and blocked in 2% BSA in
PBS, at room temperature for 2 h. For anti-cardiolipin ELISA Immulon 2HB plates
(Thermo Fisher) were coated with 75 Î¼g/ml of cardiolipin dissolved in
100% ethanol at 25Â°C overnight. Sera were diluted 1:200 and incubated on
coated plates at 25Â°C for 2 h. Plates were then incubated with
horseradish peroxidase-labeled goat anti-mouse IgG, IgG1 or IgG2c Fc antibody
for 1 h (eBioscience). Anti-ssDNA and anti-nRNP IgG ELISAs were obtained from
Alpha Diagnostic International. OD₄₅₀ was measured on a microplate
reader. ANAs were detected on Hep-2 slides (MBL international) at a 1:200
dilution using Alexa Flour 488-conjugated anti-mouse IgG (Jackson ImmunoResearch
Laboratories). Fluorescent intensity was semi-quantitated as previously
described^{52 (link)}.

Manni M., Gupta S., Ricker E., Chinenov Y., Park S.H., Shi M., Pannellini T., Jessberger R., Ivashkiv L, & Pernis A.B. (2018). Regulation of Age-associated B cells by IRF5 in systemic autoimmunity. Nature immunology, 19(4), 407-419.

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Antibody Binding ELISA Protocol

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To assess antibody binding, soluble protein was plated on Immulon 2HB plates (Thermo Fisher Scientific) at 2 μg/mL overnight at 4°C. In cases where capture ELISA was used, plates were pre-incubated for 2 h at room temperature (RT) with 5 μg/mL GNA lectin (Sigma) or 2 μg/mL anti-AviTag (Genscript) and washed 3× with PBS+ 0.05% Tween 20 (PBS-T) before antigen plating overnight. Between each of the subsequent incubation steps, plates were washed 3× with PBS-T. Non-specific binding was blocked by incubation with 5% fetal bovine serum (FBS) (Gibco) diluted in PBS-T for 1 h at RT. Primary monoclonal antibodies were diluted in 5% FBS-PBST starting at 20 μg/mL with a serial 1:5 dilution (unless otherwise specified) and then added to the plate for 1 h at RT. Secondary antibody, either goat anti-human IgG (Southern Biotech) or goat anti-human IgA (Invitrogen), was diluted 1:10,000 in 5% FBS diluted in PBS-T and added for 1 h at RT. Reaction was developed by 10 min incubation with One Step Ultra-TMB (Thermo Fisher Scientific) and stopped with 1N sulfuric acid. Plate absorbances were read at 450 nm (Biotek). Data are represented as mean ± SEM for one ELISA experiment performed in duplicate. ELISA experiments were repeated with at least 2 different antibody preparation aliquots. The area under the curve (AUC) was calculated using GraphPad Prism 8.0.0.

Pilewski K.A., Wall S., Richardson S.I., Manamela N.P., Clark K., Hermanus T., Binshtein E., Venkat R., Sautto G.A., Kramer K.J., Shiakolas A.R., Setliff I., Salas J., Mapengo R.E., Suryadevara N., Brannon J.R., Beebout C.J., Parks R., Raju N., Frumento N., Walker L.M., Fechter E.F., Qin J.S., Murji A.A., Janowska K., Thakur B., Lindenberger J., May A.J., Huang X., Sammour S., Acharya P., Carnahan R.H., Ross T.M., Haynes B.F., Hadjifrangiskou M., Crowe JE J.r., Bailey J.R., Kalams S., Morris L, & Georgiev I.S. (2023). Functional HIV-1/HCV cross-reactive antibodies isolated from a chronically co-infected donor. Cell reports, 42(2), 112044.

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Collagen Film Crosslinking Protocol

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First, slurries of insoluble collagen type I (derived from bovine tendon Col (S) or skin Col(D)) were produced by swelling at 0.5% (w/v) in 50 mM acetic acid at 4 °C overnight and then homogenising on ice for 30 min at 13500 rpm using an Ultra-Turrax VD125 (VWR International Ltd., UK). Air bubbles were removed from slurries by centrifuging at 2500 rpm for 5 min (Hermle Z300, Labortechnik, Germany). Collagen films of ∼8 µm thickness were prepared by pipetting 100 μL/well of these slurries directly in Immulon 2HB plates (Thermo Scientific) and drying for 48 h in a laminar flow cabinet.
Films were cross-linked (XL) with carbodiimide (EDC) in combination with succinimide (NHS). An EDC concentration of 11.5 mg/ml, with a molar ratio of EDC/NHS/COO^-(Col) = 5/2/1 in 95% (v/v) ethanol, was taken as standard (100%) and was varied from 1 to 200%. These crosslinking conditions were selected on the basis of our previous work where the effect of the reducing of standard crosslinking concentration down to very low levels (up to 1% EDC) on different relevant material and some cell-interactive properties was elucidated [24] (link), [32] (link). After reaction for 2 h at room temperature, the films were washed thoroughly in deionised water (15 min × 5) and dried in a laminar flow cabinet.

Davidenko N., Hamaia S., Bax D.V., Malcor J.D., Schuster C.F., Gullberg D., Farndale R.W., Best S.M, & Cameron R.E. (2018). Selecting the correct cellular model for assessing of the biological response of collagen-based biomaterials. Acta Biomaterialia, 65, 88-101.

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