Mouse MCSF (Macrophage Colony Stimulating Factor 1) ELISA Kit (MOES01724)

SKU:
MOES01724
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
P07141
Sensitivity:
9.38pg/mL
Range:
15.63-1000pg/mL
ELISA Type:
Sandwich
Synonyms:
M-CSF, CSF1, CSF-1
Reactivity:
Mouse
Tested Sample Types:
Serum, plasma and other biological fluids
€549

Description

ELISA Kit Technical ManualMSDS

Mouse MCSF (Macrophage Colony Stimulating Factor 1) 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 Mouse MCSF . 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 Mouse MCSF 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 Mouse MCSF, 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 Mouse MCSF. The concentration of Mouse MCSF in samples can be calculated by comparing the OD of the samples to the standard curve.

Assay typeSandwich
Format96T
Assay time4.5h
ReactivityMouse
Detection MethodColormetric
Detection Range15.63-1000 pg/mL
Sensitivity9.38 pg/mL
Sample Volume Required Per Well100uL
Sample TypeSerum, plasma and other biological fluids

Specificity

This kit recognizes Mouse MCSF in samples. No significant cross-reactivity or interference between Mouse MCSF 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
(pg/mL)
O.D Average Corrected
10002.468
2.514
2.4912.421
5001.576
1.586
1.5811.511
2500.947
0.941
0.9440.874
1250.478
0.5
0.4890.419
62.50.246
0.23
0.2380.168
31.250.186
0.156
0.1710.101
15.630.113
0.131
0.1220.052
00.062
0.078
0.07--

Precision

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

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

Intra-assay Precision Inter-assay Precision
Sample123123
n202020202020
Mean (pg/mL)50.7086.24389.2954.4186.56399.03
Standard deviation3.463.9416.353.614.9916.32
C V (%)6.824.574.206.635.764.09

Recovery

The recovery of Mouse MCSF 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)92-105100
EDTA plasma (n=5)87-9993
Cell culture media (n=5)95-111102

Linearity

Samples were spiked with high concentrations of Mouse MCSF 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 (%)90-10488-10394-107
Average (%)9694101
1:4Range (%)90-10382-9484-96
Average (%)968888
1:8Range (%)92-10785-9885-97
Average (%)989190
1:16Range (%)89-10284-9781-93
Average (%)969088

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

Mouse MCSF (Macrophage Colony Stimulating Factor 1) ELISA Kit (MOES01724) 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 100µl of standard solutions into the standard wells.
    3. Add 100µl of Sample / Standard dilution buffer into the control (zero) well.
    4. Add 100µl 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 100µL 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 350µL 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 100µL 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 90µL 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.

Mouse MCSF (Macrophage Colony Stimulating Factor 1) ELISA Kit (MOES01724) Protein Information

UniProt Protein Function:M-CSF: Cytokine that plays an essential role in the regulation of survival, proliferation and differentiation of hematopoietic precursor cells, especially mononuclear phagocytes, such as macrophages and monocytes. Promotes the release of proinflammatory chemokines, and thereby plays an important role in innate immunity and in inflammatory processes. Plays an important role in the regulation of osteoclast proliferation and differentiation, the regulation of bone resorption, and is required for normal bone development. Required for normal male and female fertility. Promotes reorganization of the actin cytoskeleton, regulates formation of membrane ruffles, cell adhesion and cell migration. Plays a role in lipoprotein clearance. Aberrant expression of CSF1 or CSF1R can promote cancer cell proliferation, invasion and formation of metastases. Overexpression of CSF1 or CSF1R is observed in a significant percentage of breast, ovarian, prostate, and endometrial cancers. Aberrant expression of CSF1 or CSF1R may play a role in inflammatory diseases, such as rheumatoid arthritis, glomerulonephritis, atherosclerosis, and allograft rejection. 3 isoforms of the human protein are produced by alternative splicing.
UniProt Protein Details:

Protein type:Membrane protein, integral; Cytokine

Cellular Component: extracellular space; membrane; perinuclear region of cytoplasm; extracellular region; integral to membrane; plasma membrane; receptor complex

Molecular Function:protein binding; protein homodimerization activity; growth factor activity; cytokine activity; macrophage colony stimulating factor receptor binding

Biological Process: positive regulation of monocyte differentiation; ossification; macrophage differentiation; immune system process; positive regulation of osteoclast differentiation; positive regulation of cellular protein metabolic process; reproductive developmental process; positive regulation of odontogenesis of dentine-containing teeth; positive regulation of multicellular organism growth; osteoclast differentiation; odontogenesis; cell proliferation; positive regulation of mononuclear cell proliferation; homeostasis of number of cells within a tissue; positive regulation of protein kinase activity; positive regulation of cell proliferation; innate immune response; positive regulation of Ras protein signal transduction; regulation of ossification; inflammatory response; positive regulation of macrophage differentiation; positive regulation of cell-matrix adhesion; positive regulation of cell migration

UniProt Code:P07141
NCBI GenInfo Identifier:1345843
NCBI Gene ID:12977
NCBI Accession:P07141.2
UniProt Secondary Accession:P07141,Q3U395, Q8R3C8,
UniProt Related Accession:P07141
Molecular Weight:29,353 Da
NCBI Full Name:Macrophage colony-stimulating factor 1
NCBI Synonym Full Names:colony stimulating factor 1 (macrophage)
NCBI Official Symbol:Csf1
NCBI Official Synonym Symbols:op; Csfm; MCSF; C87615
NCBI Protein Information:macrophage colony-stimulating factor 1; osteopetrosis
UniProt Protein Name:Macrophage colony-stimulating factor 1
Protein Family:C-C motif chemokine
UniProt Gene Name:Csf1
UniProt Entry Name:CSF1_MOUSE
View AllClose