KRIBIOLISA™ SP 2/0 HCP ELISA

SKU: KBBP05 Category:

₹35,000.00

Immunoassay for the quantitative measurement of SP 2/0 Host Cell Proteins as a contaminant in various biological preparations.


Availability: Backorder | Pack Size: 1 x 96 wells

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Description

Introduction:
Recombinant expression by the hybridoma cell line SP2/0 is a relatively simple and cost effective method for production of monoclonal antibodies intended for use as therapeutic agents in humans.

Intended Use:
This generic kit is intended in determining the presence of SP2/0 Host Cell Proteins contamination in various products that are manufactured through recombinant expression in SP2/0 cells. The kit has been validated successfully for testing of in process and final product HCPs in variety of products regardless of growth and purification process.

Principle:
This assay is based on the Sandwich ELISA procedure. Microwells are coated with anti- SP2/0 antibody. Addition of Anti- SP2/0:HRP Conjugate and standards or samples containing SP2/0 HCPs will form a immune complex with the coated anti- SP2/0 antibody in microwell. A washing cycle will remove free Anti- SP2/0:HRP Conjugate from the reaction well. Addition of TMB Substrate will produce blue color in microwells conataining immune complex. Addition of stop solution will terminate the enzymatic reaction and blue color will be converted to yellow color which is directly proportional to the concentration of SP2/0 HCPs present and absorbance value is measured at 450nm.










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Additional information

Sample Type

Cell Culture Supernatant and Other Biological Preparations
Calibration Range

0 ng/ml – 300 ng/ml
Sensitivity

1 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:
Note: ALL STEPS MUST BE PERFORMED AT 37?C
Bring all reagents to room temperature prior to use. It is strongly recommended that all Standards and Samples be run in duplicates. A standard curve is required for each assay.
1. Pipette out 50?l of Anti- SP2/0:HRP Conjugate into each well.
2. Pipette out 100?l of Standards and Samples into the respective wells as mentioned in the work list.
3. Cover the plate and incubate it for 90 minutes at 37?C.
4. Aspirate and wash plate 5 times with Wash Buffer (1X) and blot residual buffer by firmly tapping plate upside down on absorbent paper. Wipe off any liquid from the bottom outside of the microtiter wells as any residue can interfere in the reading step. All the washes should be performed similarly.
5. Add 100?l of TMB Substrate solution and incubate in the dark for 30 minutes at 37?C. Monitor the color development at every 10 minutes.
6. Pipette out 100?l of Stop Solution. Wells should turn from blue to yellow in color.
7. Read the absorbance at 450 nm blanking on the zero standard.

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