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Product Summary	The Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit,This kit is a solid phase-phase sandwich enzyme linked immuno sorbent assay (ELISA). Samples, including standards of known target factor concentrations and unknowns are pipetted into these wells. During the first incubation, the target factor antigen and a biotinylated monoclonal antibody specific for target protein are simultaneously incubated. After washing, the enzyme (streptavidin-peroxydase) is added. After incubation and washing to remove the entire unbound enzyme, a substrate solution which is acting on the bound enzyme is added to induce a colored reaction product. The intensity of this colored product is directly proportional to the concentration of target factor present in the samples.
Product name	Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit
Catalogue No.	EIA06571h
Product Datasheet	Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Download Manual
Alias	Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit
Background	Nicotinic acid adenine dinucleotide phosphate, (NAADP), is a Ca²⁺-mobilizing second messenger synthesised in response to extracellular stimuli. Like its mechanistic cousins, IP3 and cyclic adenosine diphosphoribose (Cyclic ADP-ribose), NAADP binds to and opens Ca²⁺ channels on intracellular organelles, thereby increasing the intracellular Ca²⁺ concentration which, in turn, modulates sundry cellular processes (see Calcium signalling). Structurally, it is a dinucleotide that only differs from the house-keeping enzyme cofactor, NADP by a hydroxyl group (replacing the nicotinamide amino group) and yet this minor modification converts it into the most potent Ca²⁺-mobilizing second messenger yet described. NAADP acts across phyla from plants to man.
Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Data Examples
Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Recovery	Matrix	Recovery range(%)	Average(%)
	serum(n=5)	88-104	96
	EDTA plasma(n=5)	86-104	98
	heparin plasma(n=5)	85-103	96

Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Linearity	The linearity of the Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit was assayed by testing samples spiked with appropriate concentration of and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
	Sample	1:2	1:4	1:8	1:16
	serum(n=5)	95-105%	85-101%	91-104%	95-102%
	EDTA plasma(n=5)	84-97%	91-99%	82-99%	85-96%
	heparin plasma(n=5)	85-99%	80-94%	81-91%	85-98%

Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Assay Precision
	Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Intra-Assay Precision			Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Inter-Assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	40	40	40
Mean	29.1	123	531	32.8	128	495
CV%	6.5	4.1	3.5	8.5	5	6.7

Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit name	96Tests	48Tests	Storage
Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit antibody precoated plate	8×12	8×6	4/-20℃
Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Standards	2vial	1vial	4/-20℃
Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Biotinylated antibody(1:100)	1vial	1vial	4/-20℃
Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Enzyme conjugate(1:100)	1vial	1vial	4/-20℃
Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Enzyme diluent	1vial	1vial	4/-20℃
Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit antibody diluent	1vial	1vial	4/-20℃
Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Standard diluent	1vial	1vial	4/-20℃
Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Sample diluent	1vial	1vial	4/-20℃
Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Washing buffer(1:25)	1vial	1vial	4/-20℃
Colour Reagent A	1vial	1vial	4/-20℃
Colour Reagent B	1vial	1vial	4/-20℃
Colour Reagent C	1vial	1vial	4/-20℃
Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Instruction	1set	1set	RT

【Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Necessary for testing their own test facilities and equipment】

1.Microplate reader(Main wavelength 450nm filter,sub wavelength 630nm filter).

2.Washer(adjustable amount of liquid injection to ensure that each well 350μl lotion without overflow).

3.Clean benches,biological safety cabinets,fume hoods.

4.High-precision single-channel dispenser(range 0.5-10μl-20μl,20-200μl,200-1000μl).

5.High-precision multi-channel plus liquid(8 or 12,the range of 50-300μl of).

6.37℃ incubator.

7.Low temperature centrifuge.

8.Refrigerators(4℃,-20℃,-86℃).

9.Analytical balance.

10.Scissors,tweezers,pliers,and so on.

11.Swirl mixing device,low-frequency oscillator,and so on.

【Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Necessary for testing their own testing supplies and reagents】

1.Centrifuge tube(capacity of 1.5ml,5ml,and so on).

2.Disposable tip(range of 0.5-10μl-20μl,20-200μl,200-1000μl).

3.Pure water or distilled water.

4.Coordinate paper.

5.Absorbent paper.

6.EDTA,sodium citrate,heparin

Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Summary of operating procedures

1.Prepare reagents,samples and standards.

2.Add the prepared sample and the standard reaction at 37℃ for 90 minutes.

3.Wash the plate ,add the biotinylated antibody working solution,37℃ for 60 minutes.

4.Wash three times to add the Enzyme working solution,37℃ for 30 minutes.

5.Wash five times,and then add the Colour Reagent solution,37℃ within 30minutes.

6.Add the Colour Reagent.

7.Microplate reader measured OD values within 30 minutes.

8.Calculate the factor content of specimens tested.

Human Nicotinic Acid Adenine Dinucleotide Phosphate (NAADP) ELISA Kit Sample preparation

Serum,The collected whole blood was allowed to stand at 4 degrees overnight or centrifuged(1000 x g)for 10 minutes. The supernatant was collected and stored at -20℃ or -80℃. Avoid repeated freeze-thaw cycles.

Plasma,Collect plasma using 5% sodium citrate(sodium Citrate:blood=1:9)as an anticoagulant. Centrifuge samples for 15 minutes at 1000×g at 2 - 8℃ within 30 minutes of collection. Plasma was immediately removed and tested,or stored at -20℃ or -80℃ after dispensing. Avoid repeated freeze-thaw cycles.

Urine,Aseptically collect the first urine of the day. Centrifuge to remove particulate matter,assay immediately or aliquot and store at ≤-20℃. Avoid repeated freeze-thaw cycles.

Tissue homogenates,The preparation of tissue homogenates will vary depending on the type of tissue. For this assay,tissue was rinsed in ice-cold PBS(0.01 mol/L,pH 7.2-7.6)to completely remove excess blood and weighed prior to homogenization. The tissue was cut into small pieces,a certain amount of PBS was added,homogenized with a high-speed homogenizer or other homogenizer. The resulting suspension was sonicated with an ultrasonic cell disruptor and the cell membrane was disrupted and centrifuged at 5000×g for 5 minutes. Immediately remove the supernatant and perform the assay,or dispense and store at ≤ -20℃.

For other types of samples,please contact us.

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