Human SAP (Serum Amyloid P Component) ELISA Kit (HUES02336)

SKU:
HUES02336
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P02743
Sensitivity:
3.75ng/mL
Range:
6.25-400ng/mL
ELISA Type:
Sandwich
Reactivity:
Human
Tested Sample Types:
Serum, plasma and other biological fluids
€539

Description

ELISA Kit Technical ManualMSDS

Human SAP (Serum Amyloid P Component) ELISA Kit

This ELISA kit uses Sandwich-ELISA as the method. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Human SAP . Standards or samples are added to the appropriate micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Human SAP and Avidin-Horseradish Peroxidase (HRP) conjugate are added to each micro plate well successively and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Human SAP, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by adding Stop Solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm 2 nm. The OD value is proportional to the concentration of Human SAP. The concentration of Human SAP in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityHuman
Detection MethodColormetric
Detection Range6.25-400 ng/mL
Sensitivity3.75 ng/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Human SAP in samples. No significant cross-reactivity or interference between Human SAP and analogues was observed.

Typical Data

As the OD values of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique, washing technique or temperature effects), the operator should establish a standard curve for each test. Typical standard curve and data is provided below for reference only.

Concentration
(ng/mL)
O.D Average Corrected
4002.377
2.429
2.4032.348
2001.507
1.563
1.5351.48
1000.887
0.851
0.8690.814
500.476
0.486
0.4810.426
250.274
0.254
0.2640.209
12.50.154
0.154
0.1540.099
6.250.103
0.109
0.1060.051
00.05
0.06
0.055--

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Human SAP were tested 20 times on one plate, respectively.

Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Human SAP were tested on 3 different plates, 20 replicates in each plate.

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (ng/mL)20.7555.89182.0221.4756.47166.58
Standard deviation1.312.918.901.432.956.36
C V (%)6.315.214.896.665.223.82

Recovery

The recovery of Human SAP spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Serum (n=5)90-10196
EDTA plasma (n=5)89-10195
Cell culture media (n=5)89-10495

Linearity

Samples were spiked with high concentrations of Human SAP and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Serum (n=5) EDTA plasma (n=5) Cell culture media (n=5)
1:2Range (%)94-10790-10486-96
Average (%)1029691
1:4Range (%)92-10584-9682-95
Average (%)999188
1:8Range (%)89-10484-9884-96
Average (%)969090
1:16Range (%)90-10680-9386-102
Average (%)978793

Kit Components & Storage

An unopened kit can be stored at 4'C for 1 month. If the kit is not used within 1 month, store the items separately according to the following conditions once the kit is received.

Item Specifications Storage
Micro ELISA Plate(Dismountable)8 wells X12 strips-20'C, 6 months
Reference Standard2 vials
Concentrated Biotinylated Detection Ab (100X)1 vial, 120 uL
Concentrated HRP Conjugate (100X)1 vial, 120 uL-20'C(shading light), 6 months
Reference Standard & Sample Diluent1 vial, 20 mL4'C, 6 months
Biotinylated Detection Ab Diluent1 vial, 14 mL
HRP Conjugate Diluent1 vial, 14 mL
Concentrated Wash Buffer (25X)1 vial, 30 mL
Substrate Reagent1 vial, 10 mL4'C(shading light)
Stop Solution1 vial, 10 mL4'C
Plate Sealer5 pieces
Product Description1 copy
Certificate of Analysis1 copy

Human SAP (Serum Amyloid P Component) ELISA Kit (HUES02336) Assay procedure

    1. Set standard, test sample and control (zero) wells on the pre-coated plate and record theirpositions. It is recommended to measure each standard and sample in duplicate. Note: addall solutions to the bottom of the plate wells while avoiding contact with the well walls. Ensuresolutions do not foam when adding to the wells.
    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 (serum, plasma, tissue homogenates and otherbiological fluids) into test sample wells.
    5. Cover the plate with the sealer provided in the kit and incubate for 90 min at 37 °C.
    6. Aspirate the liquid from each well, do not wash. Immediately add 100 µL of BiotinylatedDetection Ab working solution to each well. Cover the plate with a plate seal and gently mix.Incubate for 1 hour at 37 °C.
    7. Aspirate or decant the solution from the plate and add 350 µL of wash buffer to each welland incubate for 1-2 minutes at room temperature. Aspirate the solution from each well andclap the plate on absorbent filter paper to dry. Repeat this process 3 times. Note: a microplatewasher can be used in this step and other wash steps.
    8. Add 100 µL of HRP Conjugate working solution to each well. Cover with a plate seal andincubate for 30 min at 37 °C.
    9. Aspirate or decant the solution from each well. Repeat the wash process for five times asconducted in step 7.
    10. Add 90 µL of Substrate Reagent to each well. Cover with a new plate seal and incubate forapproximately 15 min at 37 °C. Protect the plate from light. Note: the reaction time can beshortened or extended according to the actual color change, but not by more than 30min.
    11. Add 50 µL of Stop Solution to each well. Note: Adding the stop solution should be done inthe same order as the substrate solution.
    12. Determine the optical density (OD value) of each well immediately with a microplate readerset at 450 nm.

Human SAP (Serum Amyloid P Component) ELISA Kit (HUES02336) Protein Information

UniProt Protein Function:APCS: Can interact with DNA and histones and may scavenge nuclear material released from damaged circulating cells. May also function as a calcium-dependent lectin. SAP is a precursor of amyloid component P which is found in basement membrane and associated with amyloid deposits. Belongs to the pentaxin family.
UniProt Protein Details:

Protein type:Secreted; Secreted, signal peptide; Chaperone

Chromosomal Location of Human Ortholog: 1q21-q23

Cellular Component: extracellular matrix; extracellular space; protein complex; extracellular region; nucleus

Molecular Function:complement component C1q binding; unfolded protein binding; virion binding; calcium ion binding; carbohydrate binding

Biological Process: protein folding; negative regulation of virion penetration into host cell; negative regulation of acute inflammatory response; acute-phase response; innate immune response; negative regulation of monocyte differentiation; chaperone-mediated protein complex assembly; negative regulation of viral reproduction

NCBI Summary:The protein encoded by this gene is a glycoprotein, belonging to the pentraxin family of proteins, which has a characteristic pentameric organization. These family members have considerable sequence homology which is thought to be the result of gene duplication. The binding of the encoded protein to proteins in the pathological amyloid cross-beta fold suggests its possible role as a chaperone. This protein is also thought to control the degradation of chromatin. It has been demonstrated that this protein binds to apoptotic cells at an early stage, which raises the possibility that it is involved in dealing with apoptotic cells in vivo. [provided by RefSeq, Sep 2008]
UniProt Code:P02743
NCBI GenInfo Identifier:730704
NCBI Gene ID:325
NCBI Accession:P02743.2
UniProt Related Accession:P02743
Molecular Weight:223
NCBI Full Name:Serum amyloid P-component
NCBI Synonym Full Names:amyloid P component, serum
NCBI Official Symbol:APCS
NCBI Official Synonym Symbols:SAP; PTX2; HEL-S-92n
NCBI Protein Information:serum amyloid P-component; pentaxin-related; 9.5S alpha-1-glycoprotein; epididymis secretory sperm binding protein Li 92n
UniProt Protein Name:Serum amyloid P-component
UniProt Synonym Protein Names:9.5S alpha-1-glycoproteinSerum amyloid P-component(1-203)
UniProt Gene Name:APCS
UniProt Entry Name:SAMP_HUMAN
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