KRIBIOLISA™ HEK HCP ELISA

SKU: KBBP15 Category:

35,000.00

Immunoassay for the quantitative measurement of HEK293 Host Cell Proteins as a contaminant in various biological preparations.


Availability: 3 – 4 Weeks | Pack Size: 1 x 96 wells



Description

Introduction:
The KRIBIOLISA ELISA kits are used for assessing the specific biomarker in samples analytes which may be serum, plasma and cell culture supernatant as validated with the kit. The kit employs a sandwich ELISA technique which leads to a higher specificity and increased sensitivity compared to conventional competitive ELISA kits which employ only one antibody. Double antibodies are used in this kit.

Intended Use:
The KRIBIOLISA HEK HCP ELISA kit is used as an analytical tool for quantitative determination of Human HEK HCP in serum, plasma and other biological samples.

Principle:
The method employs sandwich ELISA technique. Monoclonal antibodies are pre-coated onto microwells. Samples and standards are pipetted into microwells and Human HEK HCP present in the sample are bound by the antibodies. Biotin labeled antibody is added and followed by Streptavidin-HRP is pipetted and incubated to form a complex. After washing microwells in order to remove any non-specific binding, the substrate solution (TMB) is added to microwells and color develops proportionally to the amount of Human HEK HCP in the sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.










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Additional information

Sample Type

Serum, plasma and Other Biological Samples

Calibration Range

78.125 – 5000 ng/ml

Sensitivity

46.875 ng/ml

Detection Method

Colorimetric

ELISA Type

Direct Sandwich Assay

Regulatory Status

For Research Use Only

Storage Temperature

Store the unopened product at 2-8 Deg C. Do not use past expiration date.

Protocol

Assay Procedure:
It is strongly recommended that all Standards and Samples be run in duplicates or triplicates. A standard curve is required for each assay.
1. Add 100 ul prepared Standards and Samples to respective wells.
2. Cover the plate with a sealer and incubate for 90 minutes at 37?C.
3. Aspirate and wash plate 4 times with diluted Wash Buffer (1X) and blot residual buffer by firmly tapping plate upside down on absorbent paper. Wipe of any liquid from the bottom outside of the microtiter wells as any residue can interfere in the reading step.
4. Pipette 100 ul Biotinylated HEK HCP Antibody Working Solution to all wells.
5. Cover the plate with a sealer and incubate for 60 minutes at 37?C.
6. Aspirate and wash as per Step (4) above.
7. Pipette 100 ul Streptavidin:HRP Conjugate Working Solution to all wells. Mix well.
8. Cover the plate with a sealer and incubate for 30 minutes at 37?C.
9. Aspirate and wash as per Step (4) above.
10. Pipette 100 ul TMB Substrate in all the wells. 11. Incubate the plate at 37?C for 10 minutes. DO NOT SHAKE or else it may result in higher backgrounds and worse precision. Positive wells should turn bluish in color.
12. Pipette 100 ul of Stop Solution to all wells. The wells should turn from blue to yellow in color.
13. Read the absorbance at 450 nm with a microplate within 10-15 minutes after addition of Stop solution.