产品资料

ELISAGenie/Rat SELL (L-Selectin) CLIA Kit (RTES00366)/L-Selectin) CLIA Kit (RTES00366/RTES00366

Description
Additional Information

Description

Rat SELL (L-Selectin) CLIA Kit

This kit uses Sandwich-CLIA as the method. The micro CLIA plate provided in this kit has been pre-coated with an antibody specific to Rat SELL . Standards or samples are added to the appropriate micro CLIA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat SELL 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 Rat SELL, biotinylated detection antibody and Avidin-HRP conjugate will appear fluorescence. The Relative light unit (RLU) value is measured spectrophotometrically by the Chemiluminescence immunoassay analyzer. The RLU value is positively associated with the concentration of Rat SELL. The concentration of Rat SELL in the samples can be calculated by comparing the RLU of the samples to the standard curve.

Assay type	Sandwich
Format	96T
Assay time	4.5h
Reactivity	Rat
Detection method	Chemiluminescence
Detection range	31.25-2000 pg/mL
Sensitivity	18.75 pg/mL
Sample volume	100uL
Sample type	Serum, plasma and other biological fluids
Repeatability	CV < 15%

Specificity

This kit recognizes Rat SELLin samples. No significant cross-reactivity or interference between Rat SELLand analogues was observed.

Typical Data

As the RLU 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)	RLU	Average	Corrected
2000	5494656496	55721	55693
1000	2385124097	23974	23946
500	1108510979	11032	11004
250	52075381	5294	5266
125	27202496	2608	2580
62.5	13621260	1311	1283
31.25	637711	674	646
0	2729	28	--

Precision

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

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

	Intra-assay Precision			Inter-assay Precision
Sample	1	2	3	1	2	3
n	20	20	20	20	20	20
Mean (pg/mL)	95.70	202.04	907.56	87.65	185.59	944.58
Standard deviation	11.62	21.92	61.26	11.03	21.60	105.32
C V (%)	12.14	10.85	6.75	12.58	11.64	11.15

Recovery

The recovery of Rat SELLspiked 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)	86-101	92
EDTA plasma (n=5)	92-108	98
Cell culture media (n=5)	90-102	97

Linearity

Samples were spiked with high concentrations of Rat SELLand 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)
1:2	Range (%)	93-106	95-105	101-116
1:2	Average (%)	99	100	108
1:4	Range (%)	91-104	85-96	92-107
1:4	Average (%)	96	90	98
1:8	Range (%)	98-112	98-111	89-106
1:8	Average (%)	105	104	97
1:16	Range (%)	102-119	101-115	103-116
1:16	Average (%)	110	106	110

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 CLIA Plate(Dismountable)	8 wells X12 strips	-20"C, 6 months
Reference Standard	2 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 Diluent	1 vial, 20 mL	4"C, 6 months
Biotinylated Detection Ab Diluent	1 vial, 14 mL
HRP Conjugate Diluent	1 vial, 14 mL
Concentrated Wash Buffer (25X)	1 vial, 30 mL
Substrate Reagent A	1 vial, 5 mL	4"C (shading light)
Substrate Reagent B	1 vial, 5 mL	4"C (shading light)
Plate Sealer	5 pieces
Product Description	1 copy
Certificate of Analysis	1 copy

Rat SELL (L-Selectin) CLIA Kit(RTES00366) 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 100µL of Substrate mixture solution to each well. Cover with a new plate seal andincubate for no more than 5 min at 37°C. Protect the plate from light.

11. Determine the RLU value of each well immediately.

Rat SELL (L-Selectin) CLIA Kit(RTES00366) Protein Information

UniProt Protein Function:	Cell surface adhesion protein. Mediates the adherence of lymphocytes to endothelial cells of high endothelial venules in peripheral lymph nodes. Promotes initial tethering and rolling of leukocytes in endothelia.
NCBI Summary:	lymphocyte adhesion molecule that facilitates adhesion of lymphocytes to high endothelial venules [RGD, Feb 2006]
UniProt Code:	P30836
NCBI GenInfo Identifier:	400181
NCBI Gene ID:	29259
NCBI Accession:	P30836.1
UniProt Related Accession:	P30836
Molecular Weight:	42,441 Da
NCBI Full Name:	L-selectin
NCBI Synonym Full Names:	selectin L
NCBI Official Symbol:	Sell
NCBI Official Synonym Symbols:	A.11; LECAM-1; L-selectin
NCBI Protein Information:	L-selectin
UniProt Protein Name:	L-selectin
UniProt Synonym Protein Names:	CD62 antigen-like family member L; Leukocyte adhesion molecule 1; LAM-1; Leukocyte-endothelial cell adhesion molecule 1; LECAM1; Lymph node homing receptor; Lymphocyte antigen 22; Ly-22; Lymphocyte surface MEL-14 antigen; CD_antigen: CD62L
Protein Family:	L-selectin
UniProt Gene Name:	Sell
UniProt Entry Name:	LYAM1_RAT

View AllClose

Additional Information

Reactivity:	Rat
ELISA Type:	CLIA