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No .

E-EL-H0080c

Human GDF15(Growth Differentiation Factor 15) ELISA Kit

Item No:

E-EL-H0080c

Chinese Name:

Alias:

GDF-15, MIC-1, MIC1, NAG-1, PDF, PLAB, PTGFB, TGF-PL

Keywords:

ELISA kits

Classification:

Mouse Micro(MS) Kit

Retail Price

¥ 1650

Market Price

¥ 1650

This kit is designed for the in vitro quantitative analysis of GDF15 in Human serum, plasma or other biological fluids.

Spec
- 48T
- 96T
- 96T*5

Quantity

Inventory surplus

2997

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Describe

Use

　　This kit is designed for the in vitro quantitative analysis of GDF15 in Human serum, plasma or other biological fluids.

Detection Principle

　　This kit adopts double antibody sandwich ELISA method. Anti-Human GDF15 antibody is used to encapsulate on the enzyme labeling plate. Human GDF15 in the sample or standard will bind to the encapsulated antibody during the experiment, and the free components will be washed away. Biotinylated Anti-Human GDF15 Antibody and Horseradish Peroxidase-labeled affinity were added sequentially. The anti-human GDF15 antibody binds to human GDF15 bound to the coated antibody, biotin binds specifically to the affinity element to form an immune complex, and the free components are washed away. Colorimetric substrate (TMB) was added, TMB showed blue color catalyzed by horseradish peroxidase, and turned into yellow color after addition of termination solution. The OD value was measured at 450 nm with an enzyme marker, and there was a positive ratio between the concentration of GDF15 and the OD450 value, and the concentration of GDF15 in the sample was calculated by plotting the standard curve.

Reaction type	sandwiching method
Specification	96T
Reaction time	3.5h
Reactivity	Human
Detection method	Colormetric
Detection range	23.44—1500 pg/mL
Sensitivity	14.06 pg/mL
Sample Volume	100μL
Sample Type	Serum, plasma or other biological fluids

Idiosyncrasy

　　Detects human GDF15 in samples with no significant cross-reactivity with other related proteins.

Repeatable

　　The coefficients of variation within plates and between plates were all <10%.

Typical data

　　The OD values of the standard curve will vary due to different experimental operating conditions (e.g., operator, pipetting technique, plate washing technique and stabilization conditions, etc.). The following data and curves are for reference only, and the experimenter needs to establish the standard curve according to his/her own experiment.

(pg/mL)	O.D	Average	Corrected
1500	2.463 2.517	2.49	2.435
750	1.585 1.631	1.608	1.553
375	0.989 0.975	0.982	0.927
187.5	0.478 0.498	0.488	0.433
93.75	0.275 0.261	0.268	0.213
46.88	0.171 0.147	0.159	0.104
23.44	0.103 0.113	0.108	0.053
0	0.049 0.061	0.055	--

Precision

　　Intra-plate precision: Low, medium and high concentration samples were tested 20 times on 1 plate.
　　Inter-plate precision: low concentration, medium concentration and high concentration samples were tested 20 times on 3 plates.

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	83.30	126.61	615.36	76.84	120.76	573.31
Standard deviation	4.87	6.79	32.00	4.78	6.96	27.46
C V (%)	5.85	5.36	5.20	6.22	5.76	4.79

Recovery rate

　　A known concentration of the target protein was added to five different samples and recovery experiments were done to obtain a range of recoveries and an average recovery.

Sample Type	Range (%)	Average Recovery (%)
Serum (n=8)	91-103	96
EDTA plasma (n=8)	87-102	94
Cell culture media (n=8)	88-100	95

Linearity

　　A recovery experiment was performed by adding known concentrations of target protein to 5 samples to obtain a range of recoveries and average recoveries. Dilute the 5 samples 2x, 4x, 8x and 16x to obtain the recovery range and average recovery.

		Serum (n=5)	EDTA plasma (n=5)	Cell culture media (n=5)
1:2	Range (%)	93-108	84-99	96-113
1:2	Average (%)	100	91	103
1:4	Range (%)	91-103	86-98	84-99
1:4	Average (%)	96	91	90
1:8	Range (%)	90-106	81-92	82-93
1:8	Average (%)	98	87	88
1:16	Range (%)	87-103	84-96	88-102
1:16	Average (%)	94	88	93

Kit Composition and Storage

　　Unopened kits can be stored at 2-8°C for one week; if the kit is to be used after one week, unpack the kit and store the components separately according to the conditions in the table below.

Chinese name	English name	Specification	Storage conditions
ELISA Enzyme Labeling Plate (Detachable)	Micro ELISA Plate(Dismountable)	8 holes×12 strips	-20℃, can be stored for 6 months
Lyophilized Standards	Reference Standard	2 sticks
Concentrated Biotinylated Antibody(100×)	Concentrated Biotinylated Detection Ab	1 x 120 μL
Concentrated HRP enzyme conjugate(100×)	Concentrated HRP Conjugate	1 x 120 μL	-20℃ (avoid light), can be stored for 6 months
Standard & Sample Diluent	Reference Standard & Sample Diluent	1 bottle 20 mL	2-8℃. Can be stored for 6 months
Biotinylated Antibody Diluent	Biotinylated Detection Ab Diluent	1 bottle 14 mL
Enzyme Conjugate Diluent	HRP Conjugate Diluent	1 bottle 14 mL
Concentrated Wash Solution (25×)	Concentrated Wash Buffer (25×)	1 bottle 30 mL
Substrate solution (TMB)	Substrate Reagent	1 bottle 10 mL	2-8℃(avoid light)
Reaction termination solution	Stop Solution	1 bottle 10 mL	2-8℃
Plate Lamination	Plate Sealer	5 sheets
Product manual	Product Description	1 sheet
Quality Inspection Report	Certificate of Analysis	1 sheet

　　Note: All reagent bottle caps must be screwed on tightly to prevent evaporation and microbial contamination.
Reagent volumes are based on the actual shipping version of the instructions. Reagents may be dispensed in a slightly larger volume than indicated on the label, so please measure rather than pour when using.

Self-contained items required for testing

•1. Enzyme Labeling Instrument (450nm wavelength filter)

•2. High-precision pipettes, EP tubes and disposable tips: 0.5-10 μL, 2-20 μL, 20-200 μL, 200-1000 μL

•3.37℃ thermostat, double-distilled water or deionized water

•4. Pipette

Procedure

	1. Add 100μL of standard working solution or sample into the corresponding plate wells and incubate at 37℃ for 90 minutes.
	2. Discard the liquid in the plate, immediately add 100μL of biotinylated antibody working solution and incubate at 37℃ for 60 minutes.
	3. Discard the liquid in the plate and wash the plate 3 times.
	4. add 100μL of HRP enzyme conjugate working solution to each well, incubate at 37℃ for 30 minutes, discard the liquid in the plate, and wash the plate five times.
	5. Add 90μL of substrate solution to each well and incubate at 37℃ for 15 minutes.
	6. Add 50μL of termination solution to each well.
	7. Immediately read at 450nm and process the data.