Mouse (Murine) Complement Factor H ELISA Kit

Recommended Complement Factor H Product (supplied by: Log in to see )

Antigen
Complement Factor H (CFH) ELISA Kits
  • CFH
  • im:7147412
  • si:ch211-207o17.2
  • Mud-1
  • NOM
  • Sas-1
  • Sas1
  • AHUS1
  • AMBP1
  • ARMD4
  • ARMS1
  • CFHL3
  • FH
  • FHL1
  • HF
  • HF1
  • HF2
  • HUS
  • Fh
  • fH
  • complement factor H
  • complement component factor h
  • CFH
  • cfh
  • Cfh
Epitope
AA 875-1252
Reactivity
Mouse (Murine)
Alternatives
Kits with alternative reactivity to:
17
13
10
2
2
2
2
2
2
1
1
1
Method Type
Sandwich ELISA
Detection Range
0.78-50 ng/mL
Minimum Detection Limit
0.78 ng/mL
Application
ELISA
Options
Supplier
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Catalog No. ABIN2859252
$ 369.00
Plus shipping costs $45.00
Relevance Score ABIN Method Type Sample Type Detection Range Detection Minimum Supplier References Details
12.498835 ABIN415452 Sandwich ELISA Serum, Plasma, Biological Fluids 7.8-500 ng/mL 7.8 ng/mL Log in to see
12.498835 ABIN366410 Sandwich ELISA Serum, Plasma, Tissue Homogenate 31.25-2000 ng/mL 31.25 ng/mL Log in to see
12.498835 ABIN1114336 Sandwich ELISA Serum, Plasma, Biological Fluids 15.625-1000 ng/mL 15.625 ng/mL Log in to see
1 ABIN1568959 Sandwich ELISA Biological Fluids, Plasma, Serum 15.625-1000 ng/mL 15.625 ng/mL Log in to see
1 ABIN627638 Competition ELISA Serum, Cell Culture Supernatant, Tissue Homogenate, Plasma, Body Fluids 50-1000 ng/mL 50 ng/mL Log in to see
1 ABIN955832 Sandwich ELISA 15.6-1000 ng/mL 15.6 ng/mL Log in to see
1 ABIN5520364 Sandwich ELISA Plasma, Serum, Biological Fluids, Tissue Homogenate 15.625-1000 ng/mL 15.625 ng/mL Log in to see
1 ABIN2543310 Sandwich ELISA Serum, Plasma, Biological Fluids 7.81-500 ng/mL 7.81 ng/mL Log in to see
1 ABIN4968288 Sandwich ELISA Cell Culture Supernatant, Plasma, Tissue Homogenate, Serum, Cell Lysate, Biological Fluids 15.625-1000 ng/mL 15.625 ng/mL Log in to see
1 ABIN2946627 15.625-1000 ng/mL 15.625 ng/mL Log in to see
1 ABIN454811 3.12-200 ng/mL 3.12 ng/mL Log in to see
1 ABIN5073505 Sandwich ELISA Plasma, Serum, Tissue Homogenate, Biological Fluids Log in to see
1 ABIN2112279 Competition ELISA Log in to see

General

Antigen Complement Factor H (CFH) ELISA Kits
Epitope AA 875-1252
Reactivity Mouse (Murine)
Kits with alternative reactivity to:
(17), (13), (10), (2), (2), (2), (2), (2), (2), (1), (1), (1)
Method Type Sandwich ELISA
Detection Range 0.78-50 ng/mL
Minimum Detection Limit 0.78 ng/mL
Application ELISA
Supplier Log in to see

Product Details Complement Factor H ELISA Kit

Target details Application Details Handling Images
Purpose Sandwich High Sensitivity ELISA kit for Quantitative Detection of Mouse Complement H/CFH
Brand PicoKine™
Sample Type Cell Culture Supernatant, Serum, Plasma (heparin), Plasma (EDTA)
Detection Method Colorimetric
Analytical Method Quantitative
Specificity NSO, S875-V1252
Cross-Reactivity (Details) There is no detectable cross-reactivity with other relevant proteins.
Sensitivity <20pg/mL
Material not included Microplate reader in standard size. Automated plate washer. Adjustable pipettes and pipette tips. Multichannel pipettes are recommended in the condition of large amount of samples in the detection. Clean tubes and Eppendorf tubes. Washing buffer (neutral PBS or TBS). Preparation of 0.01M TBS: Add 1.2g Tris, 8.5g Nacl
Immunogen Expression system for standard: NSO
Immunogen sequence: S875-V1252

Target details

Product Details Complement Factor H ELISA Kit Application Details Handling Images back to top
Antigen
Alternative Name CFH (CFH ELISA Kit Abstract)
Background

Protein Function: Factor H functions as a cofactor in the inactivation of C3b by factor I and also increases the rate of dissociation of the C3bBb complex (C3 convertase) and the (C3b)NBB complex (C5 convertase) in the alternative complement pathway. .

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.

Synonyms: Complement factor H,Protein beta-1-H,Cfh,Hf1,

Full Gene Name: Complement factor H

Cellular Localisation: Secreted.
Gene ID 12628
UniProt P06909
Pathways Complement System, Cellular Response to Molecule of Bacterial Origin

Application Details

Product Details Complement Factor H ELISA Kit Target details Handling Images back to top
Application Notes 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.
Comment

Tissue Specificity: Expressed by the liver and secreted in plasma.

Plate Pre-coated
Protocol mouse CFH ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for CFH has been precoated onto 96-well plates. Standards (NSO, S875-V1252) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for CFH 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 mouse CFH amount of sample captured in plate.
Assay Procedure Aliquot 0.1 mL per well of the 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL, 3.12 ng/mL, 1.56 ng/mL, 0.78 ng/mL mouse CFH standard solutions into the precoated 96-well plate. Add 0.1 mL of the sample diluent buffer into the control well (Zero well). Add 0.1 mL of each properly diluted sample of mouse cell culture supernates, serum or plasma(heparin, EDTA) to each empty well. See "Sample Dilution Guideline" above for details. We recommend that each mouse CFH standard solution and each sample is measured in duplicate.
Assay Precision
  • Sample 1: n=16, Mean(ng/ml): 14.5, Standard deviation: 0.710, CV(%): 4.9
  • Sample 2: n=16, Mean(ng/ml): 31.8, Standard deviation: 1.844, CV(%): 5.8
  • Sample 3: n=16, Mean(ng/ml): 41.2, Standard deviation: 2.760, CV(%): 6.7,
  • Sample 1: n=24, Mean(ng/ml): 15.5, Standard deviation: 0.837, CV(%): 5.4
  • Sample 2: n=24, Mean(ng/ml): 34.2, Standard deviation: 2.120, CV(%): 6.2
  • Sample 3: n=24, Mean(ng/ml): 42.5, Standard deviation: 3.102, CV(%): 7.3
Restrictions For Research Use only

Handling

Product Details Complement Factor H ELISA Kit Target details Application Details Images back to top
Handling Advice Avoid multiple freeze-thaw cycles.
Storage -20 °C,4 °C
Storage Comment Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles
Expiry Date 12 months

Images

Product Details Complement Factor H ELISA Kit Target details Application Details Handling back to top
Supplier Images
ELISA image for Complement Factor H (CFH) ELISA Kit (ABIN2859252) Mouse Complement H/CFH PicoKine ELISA Kit standard curve
'Independent Validation' Badge
Antigen Commplement Factor H
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.
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.

Validation Images
ELISA validation image for Complement Factor H (CFH) ELISA Kit (ABIN2859252) A OD450 measurements for the diluted standard supplied with kit ABIN28592...