NHP Immunogenicity ELISA Sampler Pack (SKGE0060)

.pack-tabs{margin:8px 0 4px 0;position:relative}.pack-tabs .tab-radio{position:absolute;left:-9999px;opacity:0}.pack-tabs .tab-labels{display:flex;gap:2px;border-bottom:3px solid #0066a3;margin-bottom:0;flex-wrap:wrap}.pack-tabs .tab-labels label{padding:10px 22px;cursor:pointer;background:#e9eef2;color:#0066a3;border:1px solid #cfd8df;border-bottom:none;border-radius:6px 6px 0 0;font-weight:600;font-size:14px;user-select:none;transition:all .15s ease}.pack-tabs .tab-labels label:hover{background:#dde7ef}.pack-tabs .tab-pane{display:none;padding:18px;border:1px solid #0066a3;border-top:none;background:#fff;border-radius:0 0 6px 6px}.pack-tabs .tab-radio.t-sandwich:checked~.tab-labels label.l-sandwich,.pack-tabs .tab-radio.t-competitive:checked~.tab-labels label.l-competitive,.pack-tabs .tab-radio.t-indirect:checked~.tab-labels label.l-indirect,.pack-tabs .tab-radio.t-capture:checked~.tab-labels label.l-capture{background:#0066a3;color:#fff;border-color:#0066a3}.pack-tabs .tab-radio.t-sandwich:checked~.tab-pane.p-sandwich,.pack-tabs .tab-radio.t-competitive:checked~.tab-pane.p-competitive,.pack-tabs .tab-radio.t-indirect:checked~.tab-pane.p-indirect,.pack-tabs .tab-radio.t-capture:checked~.tab-pane.p-capture{display:block}.pack-tabs .tab-pane h4{margin-top:0}NHP Immunogenicity ELISA Sampler PackThe NHP Immunogenicity ELISA Sampler Pack from Assay Genie is a comprehensive set of validated ELISA kits designed to support nhp monkey ada & vaccine response research. Anti-drug antibody and immunogenicity markers for NHP biologic safety and ADA studies. Each kit in the pack delivers reliable and reproducible quantification of its target analyte across serum, plasma, and other relevant nhp biological samples.By combining these analytes in a single sampler pack, researchers can efficiently profile multiple biomarkers relevant to monkey ada & vaccine response, accelerating biomarker discovery, mechanistic studies, and translational research in nhp models. Order the NHP Immunogenicity ELISA Sampler Pack today to streamline your multi-analyte nhp workflow.OverviewNHP Immunogenicity ELISA Sampler PackThe NHP Immunogenicity ELISA Sampler Pack from Assay Genie allows researchers to begin their investigation into nhp monkey ada & vaccine response via analysing Anti-KLH IgM, Anti-KLH IgG, Anti-KLH IgA, Total IgG and Total IgM in a 96 assay format per kit, thus allowing researchers to identify and validate analytes within the pathway.Pack ContentsSizeReactivitySensitivityRangeELISA TypeDatasheetMSDSMonkey Anti-KLH IgM (anti Keyhole Limpet Hemocyanin IgM) ELISA Kit48 AssaysMonkey0.938ng/ml1.563-100ng/mlIndirect ELISADatasheetMSDSMonkey Anti-KLH IgG (anti Keyhole Limpet Hemocyanin IgG) ELISA Kit48 AssaysMonkey0.375ng/ml0.625-40ng/mlIndirect ELISADatasheetMSDSMonkey Anti-KLH IgA (anti Keyhole Limpet Hemocyanin IgA) ELISA Kit48 AssaysMonkey0.469ng/ml0.781-50ng/mlIndirect ELISADatasheetMSDSMonkey IgG (Immunoglobulin G) ColorStep ELISA Kit48 AssaysNHPRefer to datasheetRefer to datasheetSandwich ELISADatasheetMSDSMonkey EPO-IgM (Erythropoietin Immunoglobulin M) ELISA Kit48 AssaysNHPRefer to datasheetRefer to datasheetCapture ELISADatasheetMSDSKit ComponentsThis Sampler Pack contains Sandwich and Indirect and Capture ELISA kits. Use the tabs below to view the components included with each kit type.Sandwich ELISA KitIndirect ELISA KitCapture ELISA Kit Component Quantity Storage ELISA Microplate (Dismountable) 4-12 strips 4°C for 6 months Lyophilized Standard 1 4°C/-20°C Sample/Standard Dilution Buffer 10ml 4°C Biotin-labeled Antibody(Concentrated) 60ul 4°C (Protect from light) Antibody Dilution Buffer 5ml 4°C HRP-Streptavidin Conjugate(SABC) 60ul 4°C (Protect from light) SABC Dilution Buffer 5ml 4°C TMB Substrate 10ml 4°C (Protect from light) Stop Solution 5ml 4°C Wash Buffer(25X) 15ml 4°C Plate Sealer 3 - Other materials and equipment required: Microplate reader with 450 nm wavelength filter Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips Incubator Deionized or distilled water Absorbent paper Buffer resevoir Component Quantity Storage Pre-coated ELISA Microplate (with target antigen) 96 wells (12 strips) 4°C for 6 months Positive Control 1 vial, 100 µL 4°C Negative Control 1 vial, 100 µL 4°C Sample Dilution Buffer 30 mL 4°C HRP-conjugated Anti-species IgG (Secondary Antibody) 1 vial, 12 mL (ready to use) 4°C (Protect from light) TMB Substrate 12 mL 4°C (Protect from light) Stop Solution 6 mL 4°C Wash Buffer (25X) 20 mL 4°C Plate Sealer 4 - Product Manual 1 - Other materials and equipment required: Microplate reader with 450 nm wavelength filter Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips Incubator at 37°C Deionized or distilled water Absorbent paper Buffer reservoir Component Quantity Storage Pre-coated Capture Antibody ELISA Microplate (Dismountable) 96 wells (12 strips) 4°C for 6 months Lyophilized Standard / Calibrator 2 vials 4°C / -20°C Sample / Standard Dilution Buffer 20 mL 4°C Biotinylated Detection Antibody (Concentrated) 1 vial, 120 µL 4°C (Protect from light) Detection Antibody Dilution Buffer 14 mL 4°C HRP-Streptavidin Conjugate (Concentrated) 1 vial, 120 µL 4°C (Protect from light) HRP Conjugate Dilution Buffer 14 mL 4°C TMB Substrate 10 mL 4°C (Protect from light) Stop Solution 6 mL 4°C Wash Buffer (25X) 30 mL 4°C Plate Sealer 5 - Other materials and equipment required: Microplate reader with 450 nm wavelength filter Multichannel Pipette, Pipette, microcentrifuge tubes and disposable pipette tips Incubator at 37°C Deionized or distilled water Absorbent paper Buffer reservoir Sample PreparationWhen carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types. Sample Type Protocol Serum If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. If serum separator tubes are not being used, allow samples to clot overnight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Remove serum and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Plasma Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples at 4°C for 15 mins at 1000 - g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles. Note: Over haemolysed samples are not suitable for use with this kit. Urine & Cerebrospinal Fluid Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid. Cell culture supernatant Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately. Cell lysates Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. Tissue homogenates The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenize in 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or -80°C. Tissue lysates Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C. Milk Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles. ELISA ProtocolThis Sampler Pack contains Sandwich and Indirect and Capture ELISA kits, which follow different reaction protocols. Use the tabs below to view the relevant protocol for each kit type.Sandwich ELISA ProtocolIndirect ELISA ProtocolCapture ELISA Protocol*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.Before adding to wells, equilibrate the SABC working solution and TMB substrate for at least 30 min at 37°C. When diluting samples and reagents, they must be mixed completely and evenly. It is recommended to plot a standard curve for each test.StepProtocol1.Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!2.Aliquot 0.1ml standard solutions into the standard wells.3.Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.4.Add 0.1 ml of properly diluted sample into test sample wells.5.Seal the plate with a cover and incubate at 37 °C for 90 min.6.Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.7.Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.8.Seal the plate with a cover and incubate at 37°C for 60 min.9.Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.10.Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.11.Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.12.Add 90 µl of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min.13.Add 50 µl of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.In an Indirect ELISA, the target antigen is pre-coated onto the microplate. Sample antibodies bind directly to the antigen, and an HRP-conjugated anti-species IgG secondary antibody is then used to detect bound primary antibody. Before starting, equilibrate all reagents to room temperature for at least 30 min and prepare working dilutions per the supplied datasheet.StepProtocol1.Set positive control, negative control, blank and test sample wells on the pre-coated plate. Measure controls and samples in duplicate. Record well positions.2.Add 100 µl of diluted sample, positive control or negative control into the appropriate wells. Add 100 µl of Sample Dilution Buffer to the blank well.3.Seal the plate with a cover and incubate at 37°C for 60 min.4.Remove the cover, aspirate / decant the well content, and wash the plate 4 times with 1× Wash Buffer (300 µl per well), letting the buffer soak for 1 min between washes. Pat dry on absorbent paper.5.Add 100 µl of HRP-conjugated Anti-species IgG (Secondary Antibody) working solution into each well (including the blank). Cover and incubate at 37°C for 30 min.6.Aspirate / decant and wash the plate 5 times as in step 4.7.Add 90 µl of TMB Substrate to each well. Cover and incubate at 37°C, protected from light, for 10-20 min until the positive control wells develop a clear blue colour.8.Add 50 µl of Stop Solution to each well and mix thoroughly. The colour changes from blue to yellow.9.Read the O.D. absorbance at 450 nm using a microplate reader immediately after adding the stop solution. Interpret results by comparing sample O.D. to the positive / negative control cut-off as described in the kit datasheet.*Note: Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.In a Capture (Antibody-Capture) ELISA, the microplate is pre-coated with a specific capture antibody that selectively captures the target analyte (e.g. IgM) from the sample. A biotinylated detection antibody followed by HRP-Streptavidin is then used to generate signal. Before starting, equilibrate all reagents to room temperature for at least 30 min and prepare working dilutions per the supplied datasheet.StepProtocol1.Set standard, test sample and control (zero) wells on the pre-coated capture-antibody plate. Measure each standard and sample in duplicate. Record well positions.2.Aliquot 100 µl of standard solutions into the standard wells.3.Add 100 µl of Sample / Standard Dilution Buffer into the control (zero) well.4.Add 100 µl of properly diluted sample into the test sample wells.5.Seal the plate with a cover and incubate at 37°C for 60 min.6.Aspirate / decant the well content and wash the plate 3 times with 1× Wash Buffer (300 µl per well). Pat dry on absorbent paper.7.Add 100 µl of Biotinylated Detection Antibody working solution into each well. Seal and incubate at 37°C for 60 min.8.Aspirate / decant and wash the plate 3 times as in step 6.9.Add 100 µl of HRP-Streptavidin working solution into each well. Cover and incubate at 37°C for 30 min.10.Aspirate / decant and wash the plate 5 times as in step 6.11.Add 90 µl of TMB Substrate to each well. Cover and incubate at 37°C, protected from light, for 10-20 min.12.Add 50 µl of Stop Solution to each well and mix thoroughly. The colour changes from blue to yellow.13.Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.