Complement Factor H ELISA 试剂盒
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Quick Overview for Complement Factor H ELISA 试剂盒 (ABIN2859252)
抗原
See all Complement Factor H (CFH) ELISA试剂盒抗原表位
适用
检测方法
实验类型
检测范围
应用范围
样品类型
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最低检测浓度
- 0.78 ng/mL
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原理
- Sandwich Mouse Complement Factor H/CFH ELISA Kit to quantitate Mouse Cfh in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).
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Analytical Method
- Quantitative
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特异性
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Expression system for standard: NS0
Immunogen sequence: S875-V1252Capture antibody and Detection antibody: monoclonal antibody from rat, polyclonal antibody from goat
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交叉反应 （详细）
- There is no detectable cross-reactivity with other relevant proteins.
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灵敏度
- < 20 pg/mL
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组件
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- Pre-coated 96-well strip microplate
- Standard
- Biotinylated antibody (100x)
- Avidin-Biotin-Peroxidase Complex (100x)
- Sample Diluent
- Antibody Diluent
- Avidin-Biotin-Peroxidase Diluent
- Color Developing Reagent (TMB)
- Stop Solution
- Wash Buffer (25x)
- Adhesive plate sealers
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试剂未包括
- Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional). Pipettes and pipette tips capable of precisely dispensing 0.5 μL through 1 mL volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500 mL graduated cylinders. Test tubes for dilution.
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Complement Factor H (CFH) ELISA Kit
CFH 适用: 小鼠 Colorimetric Sandwich ELISA 7.8 ng/mL - 500 ng/mL Plasma, Serum
Complement Factor H (CFH) ELISA KitCFH 适用: 人 Colorimetric Sandwich ELISA 15.6 ng/mL - 1000 ng/mL Plasma, Serum
Complement Factor H (CFH) ELISA KitCFH 适用: 大鼠 Colorimetric Competition ELISA 12.35 ng/mL - 1000 ng/mL Plasma, Serum
Complement Factor H (CFH) ELISA KitCFH 适用: 大鼠 Colorimetric Sandwich ELISA 15.6 ng/mL - 1000 ng/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Complement Factor H (CFH) ELISA KitCFH 适用: 小鼠 Colorimetric Sandwich ELISA 31.25 ng/mL - 2000 ng/mL Plasma, Serum, Tissue Homogenate
Complement Factor H (CFH) ELISA KitCFH 适用: 大鼠 Colorimetric Sandwich ELISA 15.625 ng/mL - 1000 ng/mL Plasma, Serum, Tissue Homogenate
Complement Factor H (CFH) ELISA KitCFH 适用: 人 Colorimetric Sandwich ELISA 23.438 ng/mL - 1500 ng/mL Plasma, Serum, Tissue Homogenate
Complement Factor H (CFH) ELISA KitCFH 适用: 小鼠 Colorimetric Sandwich ELISA 15.625 ng/mL - 1000 ng/mL Plasma, Serum, Tissue Homogenate
Complement Factor H (CFH) ELISA KitWide Range CFH 适用: 人 Colorimetric Sandwich ELISA 23.44 ng/mL - 1500 ng/mL Plasma, Serum
Complement Factor H (CFH) ELISA KitCFH 适用: 人 Colorimetric Sandwich ELISA 31.25-2000 ng/mL Cell Culture Supernatant, Plasma, Saliva, Serum, Tissue Homogenate, Urine
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应用备注
- Before using Kit, spin tubes and bring down all components to bottom of tube. Duplicate well assay was recommended for both standard and sample testing.
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样本量
- 100 μL
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板类型
- Pre-coated
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实验流程
- ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. An antibody has been precoated onto 96-well plates. Standards and test samples are added to the wells, a biotinylated detection antibody specific for target is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the target amount in sample captured in plate.
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实验精密度
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Intra-Assay Precision (CV%): 7.3%, 5.7%, 5.8%
Inter-Assay Precision (CV%): 7.8%, 6.8%, 7% -
限制
- 仅限研究用
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- by
- Alexander Lab, Jacobs School of Medicine and Biomedical Sciences, Department of Medicine, University of Buffalo
- No.
- #101404
- 日期
- 2017.09.20
- 抗原
- Commplement Factor H
- Lot Number
- Method validated
- ELISA
- Positive Control
- C57BL/6 mouse serum
- Negative Control
- Complement Factor H knockout mouse serum
- Notes
- Passed. ABIN2859252 detects complement factor H in C57BL/6 mouse serum samples.
- Primary Antibody
- Secondary Antibody
- Full Protocol
- Centrifuge blood obtained from mice using EDTA was, separate serum and store serum at -80°C.
- Dilute serum 1:1000 in DILUENT and add 10µl to each well. Add 90µl of diluent to each well.
- Dilute the mouse CFH standard provided with the kit twofold from 0.78-25ng/ml according to the kit manual. Add 100µl of the diluted standard to each well.
- Run both standard and samples in duplicate.
- Use diluent buffer as blank.
- Cover the plate and incubate for 90min at 37°C.
- Discard the liquid and wash the plate with 3x with PBS.
- Add 100µl biotinylated anti-mouse FH antibody working solution into each well.
- Incubate the plate for 60min at 37°C.
- Wash the plate 3x with PBS.
- Added 100µl of ABC working solution into each well and incubated at for 30min at 37°C.
- Wash plate 3x with PBS.
- Discard the liquid and dry plate by blotting.
- Add 90µl of TMB color developing agent provided with the kit and incubate the plate for 30min at 37°C.
- Added 100µl prepared TMB stop solution.
- Read OD450 after 20min using a BioRad iMark Microplate reader.
- Experimental Notes
The signal in the positive controls samples was significantly higher than the blank whereas the signal from the negative control samples was higher than the baseline.
The serum obtained from knockout mice also had absorption. Complement Factor H is a family of proteins. It is possible that the antibodies utilized for the sandwich ELISA binds to the smaller members of the family that was not deleted in the knockout. The may need more standardization with the serum since some samples from the knockout gave background.
The linear range of the kit appears to be between 0.78 and 12.5ng/ml: the highest concentration standard (25ng/ml) lies off the best fit straight line generated for the lower concentration standards (see figure B and C). The CFH concentration in the sample was therefore calculated based on the equation for the best fit straight line not taking the 25ng/ml data point into account.
生效 #101404 (ELISA)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![A OD450 measurements for the diluted standard supplied with kit ABIN2859252, blank samples, and positive and negative control samples and the corresponding calculated concentrations. B Standard curve reflecting a twofold dilution series of the kit standard between 0.78ng/ml and 12.5ng/ml. C Standard curve based on the serial standard dilutions ranging from 0.78ng/ml to 25ng/ml.]()
A OD450 measurements for the diluted standard supplied with kit ABIN2859252, blank samples, and positive and negative control samples and the corresponding calculated concentrations. B Standard curve reflecting a twofold dilution series of the kit standard between 0.78ng/ml and 12.5ng/ml. C Standard curve based on the serial standard dilutions ranging from 0.78ng/ml to 25ng/ml.
Full Methods -
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注意事项
- Avoid multiple freeze-thaw cycles.
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储存条件
- -20 °C,4 °C
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储存方法
- Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)
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有效期
- 12 months
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- Complement Factor H (CFH)
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别名
- CFH
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背景
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Background: Complement factor H (CFH), originally known as beta-1H globulin, is a serum glycoprotein that regulates the function of the alternativecomplement pathway in fluid phase and on cellular surfaces. It binds to C3b, accelerates the decay of the alternative pathway convertase C3bBb, and also acts as a cofactor for complement factor I, another C3b inhibitor. The CFH gene is located on chromosome 1q32-q32.1 within a cluster of genes encoding the regulatory complement components of the activation of C3 (RCA for 'regulators of complement activation'). This gene cluster includes decay-accelerating factor (DAF), C4-binding protein (C4BPA and C4BPB), and the factor H-related genes CFHR1, CFHR2, CFHR3, CFHR4, and CFHR5, among others. The gene family has arisen by multiple duplication events.
Gene Full Name: complement component factor h
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基因ID
- 12628
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UniProt
- P06909
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途径
- Complement System, Cellular Response to Molecule of Bacterial Origin
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