Human Inter-alpha-trypsin inhibitor heavy chain H4 / ITIH4 Antibody ELISA Kit (AEFI01103)
- SKU:
- AEFI01103
- Product Type:
- ELISA Kit
- Size:
- 96 Assays
- Reactivity:
- Human
- Detection Method:
- Indirect ELISA
Description
Product Name: | Human Inter-alpha-trypsin inhibitor heavy chain H4 / ITIH4 Antibody ELISA Kit |
SKU: | AEFI01103 |
Size: | 96T |
Reactivity: | Human |
Synonyms: | Inter-alpha-trypsin inhibitor heavy chain H4 antibody ELISA Kit, ITI heavy chain H4 antibody ELISA Kit, ITI-HC4 antibody ELISA Kit, Inter-alpha-inhibitor heavy chain 4 antibody ELISA Kit, Inter-alpha-trypsin inhibitor family heavy chain-related protein antibody ELISA Kit, IHRP antibody ELISA Kit, Plasma kallikrein sensitive glycoprotein 120 antibody ELISA Kit, Gp120, PK-120 antibody ELISA Kit, 70 kDa inter-alpha-trypsin inhibitor heavy chain H4 antibody ELISA Kit, 35 kDa inter-alpha-trypsin inhibitor heavy chain H4 antibody ELISA Kit, ITIH4 antibody ELISA Kit, IHRP antibody ELISA Kit, ITIHL1 antibody ELISA Kit, PK120 antibody ELISA Kit |
Sample Type: | Serum, Plasma, Cell Culture Supernatant, cell or tissue lysate, Other liquid samples |
Detection Method: | Indirect ELISA |
Detection Duration: | 4 hours |
Detection Wavelength: | OD450 |
Range: | 0.781-50ng/ml |
Sensitivity: | 0.469ng/ml |
Storage: | 2-8°C(Sealed), Don't cryopreserve. |
Specificity: | Specifically binds with Anti- ITIH4 , no obvious cross reaction with other analogues. |
This kit is based on indirect ELISA detection method and takes 4h assay time. The microplate provided in this kit has been precoated with antigen. Biotinylated antibody is used as detection antibody. Add standard and properly diluted sample into relevant well respectively. After incubation, wash unbound components. Add biotinylated detection antibody. Then, it binds with Anti- ITIH4 bound to precoated antigen. Wash unbound components and add HRP-Streptavidin Conjugate (SABC). Wash unbound components again and add TMB substrate solution. Then, TMB was catalyzed by HRP to produce a blue color product that turned yellow after adding a stop solution. Read the O.D. absorbance at 450nm in a microplate reader. Calculate the concentration of Anti- ITIH4 in the sample by plotting standard curve. The concentration of the target substance is proportional to the OD450 value.
Component | Size (96T) | Storage |
ELISA Microplate (Dismountable) | 8×12 | Put the rest strips into a sealed foil bag with the desiccant. Stored for 1 month at 2-8°C; Stored for 6 months at -20°C |
Lyophilized Standard | 2 vials | Put the rest standards into a desiccant bag. Stored for 1 month at 2-8°C; Stored for 6 months at -20°C |
Biotin-labeled Antibody (Concentrated, 100X) | 120 ul | 2-8°C (Avoid Direct Light) |
HRP-Streptavidin Conjugate (SABC, 100X) | 120 ul | |
TMB Substrate | 10 ml | |
Sample Dilution Buffer | 20 ml | 2-8°C |
Antibody Dilution Buffer | 10 ml | |
SABC Dilution Buffer | 10 ml | |
Stop Solution | 10 ml | |
Wash Buffer (Concentrated, 25X) | 30 ml | |
Plate Sealer | 5 pieces | |
Product Description | 1 copy |
Other materials and equipment required:
- Microplate reader (wavelength: 450nm)
- 37°C incubator (CO2 incubator for cell culture is not recommended.)
- Automated plate washer or multi-channel pipette/5ml pipettor (for manual washing purpose)
- Precision single (0.5-10μL, 5-50μL, 20-200μL, 200-1000μL) and multi-channel pipette with disposable tips (Calibration is required before use.)
- Sterile tubes and Eppendorf tubes with disposable tips
- Absorbent paper and loading slot
- Deionized or distilled water
Recovery: | Add a certain amount of Anti-ITIH4 into the sample. Calculate the recovery by comparing the measured value with the expected amount of Anti-ITIH4 in the sample. | |||||||||||||||||||||||||||||||||||
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Linearity: | Dilute the sample with a certain amount of Anti-ITIH4 at 1:2, 1:4 and 1:8 to get the recovery range. | |||||||||||||||||||||||||||||||||||
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Stability: | Perform the stability test for the sealed kit at 37°C and 2-8°C and get relevant data. | |||||||||||||||||||||||||||||||||||
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Precision(%): | Intra-assay Precision: samples with low, medium and high concentration are tested 20 times on the same plate. Inter-assay Precision: samples with low, medium and high concentration are tested 20 times on three different plates. |
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Step 1: | Add 100ul standard or sample into each well, seal the plate and static incubate for 90 minutes at 37°C. Washing: Wash the plate twice without immersion. |
Step 2: | Add 100ul biotin-labeled antibody working solution into each well, seal the plate and static incubate for 60 minutes at 37°C. Washing:Wash the plate three times and immerse for 1min each time. |
Step 3: | Add 100ul SABC working solution into each well, seal the plate and static incubate for 30 minutes at 37°C. Washing:Wash the plate five times and immerse for 1min each time. |
Step 4: | Add 90ul TMB substrate solution, seal the plate and static incubate for 10-20 minutes at 37°C. |
Step 5: | Add 50ul stop solution. Read at 450nm immediately and calculate. |
Serum: | Place whole blood sample at room temperature for 2 hours or at 2-8°C overnight. Centrifuge for 20min at 1000xg and collect the supernatant to detect immediately. Or you can aliquot the supernatant and store it at -20°C or -80°C for future’s assay. |
Plasma: | EDTA-Na2/K2 is recommended as the anticoagulant. Centrifuge samples for 15 minutes at 1000×g 2-8°C within 30 minutes after collection. Collect the supernatant to detect immediately. Or you can aliquot the supernatant and store it at -20°C or -80°C for future’s assay. For other anticoagulant types and uses, please refer to the sample preparation guideline. |
Tissue Sample: |
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Cell Culture Supernatant: | Collect the supernatant: Centrifuge at 2500 rpm at 2-8℃ for 5 minutes, then collect clarified cell culture supernatant to detect immediately. Or you can aliquot the supernatant and store it at -80°C for future’s assay. |
Cell Lysate: |
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Other Biological Sample: | Centrifuge samples for 15 minutes at 1000×g at 2-8℃. Collect the supernatant to detect immediately. Or you can aliquot the supernatant and store it at -80°C for future’s assay. |