ELISA/assay

HEK293T HCP ELISA Kit

DEIASL412

96T

HEK293T HCP ELISA Kit

HEK 293 HCP

HEK293T HCP ELISA Kit

sELISA

HEK 293 HCP

Quantitative

bulk products

96T

The product should be stored at 2-8°C while being protected from light; the shelf life is 8 months.

HEK 293 HCP

HEK 293 host cell protein assay, HCP assay, Host cell protein assay, HCP ELISA kit

The HEK293T Host Cell Protein (HCP) ELISA kit is designed to quantitatively measure HCP contamination in bulk products expressed in HEK293T expression systems.

This product is applicable for assay of 293T HCP in test samples by double-antibody sandwich method. The standard and test samples are added to reaction wells on microplate pre-coated with 293T antibody for incubation. The present 293T HCP quantitatively binds to the antibodies in the microplate, while unbound substances are removed after plate washing. Add Anti 293T HCP -Biotin and Streptavidin HRP successively to form an antibody - antigen - biotinylated antibody - enzyme-labeled avidin complex. The protein content in the test sample can be indicated by the color development degree of TMB. Please read the instructions for use carefully and check the components of the kit before use.

This kit is designed for the quantitative detection of HCP (host cell protein) content in biopharmaceuticals expressed on 293T cells by using a double-antibody sandwich method, and can be used to detect all components of HCP (host cell protein) in 293T cell.

1. 293T HCP Coated Plate. 8 wells × 12 strips × 1 piece. Ready-to-use 2. Anti-293T HCP-Biotin (detection antibody). 150 μL × 1 vial. 1: 99, dilute with Diluent Buffer 3. Streptavidin HRP (enzyme conjugate). 375 μL × 3 vials. 1: 9, dilute with Diluent Buffer 4. 293T HCP Standard (standard). 600 μL × 1 vial (81 μg/mL). Operate as per the recommended dilution procedure 5. Diluent Buffer. 1 g× 1 vial. Use 1×PBS-T for current use 6. 10× PBS-T Wash Buffer. 50 mL× 1 bottle. 1: 9, dilute with deionized water 7. TMB Substrate. 15 mL× 1 bottle. Ready-to-use 8. Stop Solution. 15 mL× 1 bottle. Ready-to-use 9. Plate Sealer. 1 piece. Ready-to-use 10. Instructions for Use. 1 copy. Ready-to-use

1. Plate reader 2. Thermostat plate shaker 3. Micro pipette and tips 4. Deionized water 5. Unused filter paper 6. Vortex shaker

1. Temperature equilibration: Transfer reagents to be used to room temperature (18-25°C) environment and equilibrate the temperature for 30 minutes. 2. Preparation: a. 1× PBS-T Wash Buffer: Calculate the volume of working buffer required, measure an appropriate amount of 10× PBS-T Wash Buffer, dilute with deionized water at 1:9, and mix well for later use. b. Working solution of Diluent Buffer: Take an appropriate amount of diluent dry powder according to the experimental requirements, and dissolve it with 1× PBS-T at a ratio of 1g/100mL. c. Detection antibody working buffer: Calculate the volume of working buffer required, measure an appropriate amount of biotinylated antibody, dilute with Diluent Buffer at 1:99, and mix well for later use. d. Enzyme conjugate working buffer: Calculate the volume of working buffer required, measure an appropriate amount of enzyme conjugate, dilute with Diluent Buffer at 1:9, and mix well for later use. e. The standard and test samples should be diluted with the Diluent Buffer. 3. Dilution of standard:

1. Mix all reagents well before use to avoid bubbles. 2. Confirm the number of stripe plates required based on the number of experimental wells. Put remaining strip plates back to aluminum foil bags with desiccants and seal the bag. 3. Loading: Add standard, sample, and negative control into respective wells at 100 μL/well. Seal the plate with a plate sealer, and allow to react for 1.5 h at room temperature. 4. Plate washing: Discard the liquid in each well, and fill the wells with 1× PBS-T Wash Buffer (250 μL/well). Stand for 30 seconds and discard the liquid in each well. Repeat the procedure for 3 times, and pat the plate dry on the filter paper after each washing. 5. Addition of biotinylated detection antibody working buffer: Add 100 μL of biotinylated detection antibody working buffer to each well, and react for 45 minutes at room temperature after sealing the plate with a plate sealer. 6. Plate washing: Discard the liquid in each well, and fill the wells with 1× PBS-T Wash Buffer (250 μL/well). Stand for 30 seconds and discard the liquid in each well. Repeat the procedure for 3 times, and pat the plate dry on tissue after each washing. 7. Addition of enzyme conjugate working buffer: Add 100 μL of enzyme conjugate working buffer to each well, and react for 30 minutes at room temperature after sealing the plate with a plate sealer. 8. Plate washing: Discard the liquid in each well, and fill the wells with 1× PBS-T Wash Buffer (250 μL/well). Stand for 2 minutes and discard the liquid in each well. Repeat the procedure for 3 times, and pat the plate dry on tissue after each washing. 9. Color development: Add 100 μL of TMB Substrate to each well, gently shake to mix well, seal the plate with a plate sealer, and place the plate at 25°C for 15 minutes for color development reaction. 10. Assay: Add 100 μL of Stop Solution to each well and gently shake to mix well. Measure the optical density (OD) value of each well with a microplate reader at a primary wavelength of 450 nm.

The 4-parameter fitting method is recommended for the linear fitting and calculation of the product. OD processing of the standard curve (the following example is provided as reference only, and the results from actual detection shall prevail): The standard curve is obtained by 4-parameter fitting with the theoretical standard concentrations and the corresponding OD values (as shown in the figure below)

CV%≤10%, RE%≤±15%

Assay range: 37-27000 ng/mL Limit of quantification: 37 ng/mL

1. The reagents should be stored as indicated on the label, and should be equilibrated to room temperature before use. 2. Before using the pre-coated strip plates, please equilibrate to room temperature before opening the secondary packaging. Strip plates not used in the experiment should be put back to the package immediately, and the package should be sealed tight. The plates may be stored for 1 month at 4°C. Other unused reagents should be packaged or covered. 3. The volumes of standard, biotin, and enzyme conjugate are all very small. Please perform rapid centrifugation before use to let liquid on the tube wall or cap gather at tube bottom. 4. Please use disposable tips during experimental operation to avoid cross contamination. 5. Please check each reagent in the kit before use. To obtain accurate assay results, it is of special importance to mix well or shake well the reagents for dilution, loading, and reaction termination. 6. When washing residual Wash Buffer in the reaction wells, pat the plate dry adequately on clean tissue papers until watermark is no longer visible. Do not put the tissue paper into the well to absorb the liquid. 7. The TMB Substrate is photosensitive, thus long-time exposure to illumination should be avoided; avoid contact with metal, otherwise, the assay results may be affected. 8. The kit is for single use. Please use within the shelf life.

The reagent is for assay of 293T HCP in test samples only.

The HEK293T Host Cell Protein (HCP) ELISA kit is designed to quantitatively measure HCP contamination in bulk products expressed in HEK293T expression systems.

No. Components Size Storage Conditions 1 293T HCP Coated Plate 8 wells × 12 strips × 1 piece 2-8°C 2 Anti-293T HCP-Biotin (detection antibody) 150 μL × 1 vial 2-8°C 3 Streptavidin HRP (enzyme conjugate) 375 μL × 3 vials 2-8°C 4 293T HCP Standard (standard) 600 μL × 1 vial (81 μg/mL) 2-8°C 5 Diluent Buffer 1 g× 1 vial 2-8°C 6 10× PBS-T Wash Buffer 50 mL× 1 bottle 2-8°C 7 TMB Substrate 15 mL× 1 bottle 2-8°C 8 Stop Solution 15 mL× 1 bottle 2-8°C 9 Plate Sealer 1 piece 2-8°C 10 Instructions for Use 1 copy 2-8°C

See the individual product datasheet

See the individual product datasheet

See the individual product datasheet

See the individual product datasheet

See the individual product datasheet