Name: Rat phosphor（P）ELISA Kit
Brand: Biological test kits, ELISA (enzyme-linked immunosorbent assay) kits, chemical reagents-Shanghai IDEAL MEDICAL TECH Co. Ltd.
SKU: ADL11219R1
Availability: InStock
Rating: 5 (1 reviews)

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ELISA Kit

Researchuse ELISA kit

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Home > ELISA Kit > Researchuse ELISA kit > Rat ELISA kit

Description

Rat phosphor（P）ELISA Kit
Specifications	48T/96T
article number	ADL11219R1
Species	rat
Sample type	serum, plasma, tissue homogenate,cell lysate, cell Culture supernatant etc.
Detection range	See the manual for details
Sensitivity	See the manual for details
Response time	1-5h
Sample volume	10ul
Detection wavelength	450nm
Delivery date	3-5 days
Manual	Consult our salesperson for obtaining

Principle of the assay

The kit assay Rat P level in the sample，use Purified Rat P antibody to coat microtiter plate wells, make solid-phase antibody, then add P to wells, Combined P antibody which With HRP labeled, become antibody - antigen - enzyme-antibody complex, after washing Completely, Add TMB substrate solution,TMB substrate becomes blue color At HRP enzyme-catalyzed, reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm. The concentration of Rat P in the samples is then determined by comparing the O.D. of the samples

to the standard curve.

Kit composition

Serial number	English name	specification
1	Wash solution(30X)	20ml×1 bottle
2	HRP-Conjugate reagent	6ml×1 bottle
3	Microelisa stripplate	12 holes×8 strips
4	Sample diluent	6ml×1 bottle
5	Chromogen Solution A	6ml×1 bottle
6	Chromogen Solution B	6ml×1/bottle
7	Stop solution	6ml×1 bottle
8	Standard	0.5ml×1 bottle
9	Standard diluent	1.5ml×1 bottle
10	Product Description	1 serving
11	Sealing plate membrane	2 sheets
12	Sealing bag	1 piece

Note: Please check whether the label and quantity of the reagents in the kit are consistent with the table before use.

Specimen requirements

1. extract as soon as possible after Specimen collection,and according to the relevant literature, and should be experiment as soon as possible after the extraction. If it can’t, specimen can be kept in -20 ℃ to preserve, Avoid repeated freeze-thaw cycles.
2. Can’t detect the sample which contain NaN3, because NaN3 inhibits HRP active.

Assay procedure

1. Dilute and add sample:Dilute Original density Standard as follow table:

400pg/ml	5 Standard	150ul Original density Standard+150ul Standard diluent
200pg/ml	4 Standard	150ul 5 Standard+150ul Standard diluent
100pg/ml	3 Standard	1 50ul 4 Standard+150ul Standard diluent
50pg/ml	2 Standard	150ul 3 Standard +150ul Standard diluent
25pg/ml	1 Standard	150ul 2 Standard +150ul Standard diluent

2.add sample： Set blank wells separately (blank comparison wells don’t add sample and

HRP-Conjugate reagent, other each step operation is same). testing sample well.Add 50μl of

standard to Microelisa stripplate, add Sample dilution 40μl to testing sample well, then add

testing sample 10μl (sample final dilution is 5-fold), add sample to wells , don’t touch the well

wall as far as possible, and Gently mix.

3.Incubate: After closing plate with Closure plate membrane ,incubate for 30 min at 37℃.

4.Configurate liquid: 30-fold(or 20-fold) wash solution diluted 30-fold (or 20-fold) with distilled

water and reserve.

5.washing：Uncover Closure plate membrane, discard Liquid, dry by swing, add washing buffer

to every well, still for 30s then drain, repeat 5 times, dry by pat.

6.add enzyme：Add HRP-Conjugate reagent 50μl to each well, except blank well.

7.incubate：Operation with 3.

8.washing：Operation with 5.

9.color：Add Chromogen Solution A 50ul and Chromogen Solution B 50ul to each well, evade

the light preservation for 10 min at 37℃

10.Stop the reaction：Add Stop Solution50μl to each well, Stop the reaction(the blue color

change to yellow color).

11.assay：take blank well as zero , Read absorbance at 450nm after Adding Stop Solution and

within 15min.

Important notes

1. The kit takes out from the refrigeration environment should be balanced 15-30 minutes in

the room temperature, ELISA plates coated if has not use up after opened, the plate should

be stored in Sealed bag.

2. washing buffer will Crystallization separation, it can be heated the water helps dissolve

when dilute . Washing does not affect the result.

3. add Sample with sampler Each step, And proofread its accuracy frequently, avoids the

experimental error. add sample within 5 min, if the number of sample is much , recommend

to use Volley .

4. if the testing material content is excessively higher (The sample OD is bigger than the first

standard well ),please dilute Sample (n-fold), Please diluente and multiplied by the dilution

factor.（×n×5）.

5. Closure plate membrane only limits the disposable use, to avoid cross-contamination.

6. The substrate evade the light preservation.

7. Please according to use instruction strictly, The test result determination must take the

microtiter plate reader as a standard.

8. All samples, washing buffer and each kind of reject should according to infective material

process.

9. Do not mix reagents with those from other lots

Storage conditions and validity period
1. Store at 2-8℃, do not freeze, the validity period is 6 months.
2. After opening and use, put the coated microplate into a ziplock bag with desiccant, seal the ziplock bag, and put all reagents back into the refrigerator at 2-8°C.

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