Host Cell Proteins Detection Kit, MDCK system (CAT#: HCP-LX013)

96 reactions

Anti-MDCK:HRP Conjugate;
MDCK HCP Standards Set, A-H;
TBS Wash Concentrate, 20X;
Stop Solution;
TMB Substrate for ELISA;
Anti-MDCK Coated Microtiter Strips;

Mammalian

MDCK

Dog

The detection kit is based off of "sandwich"ELISA methods. Host Cell Protein antibodies are immobilized to 96 well plates. Samples are added, and Host Cell Proteins are bound by the antibodies. Biotinylated Host Cell Protein antibody is then added, followed by strepavidin HRP, which binds to biotin. TMB substrate is then added, and HRP catalyzes the conversion of TMB to a colored product. The reaction is then stopped and read on a standard colorimetric plate reader.

96-well plate

Store at 4°C.

Sodium azide will interfere with this assay and should not be used in samples or buffers.

Gel Packs

1. Pipette 100uL of anti-MDCK:HRP into each well.
2. Pipette 50uL of standards, controls and samples into wells indicated on work list.
3. Cover & incubate on orbital shaker at 400-600rpm for 3 hours at room temperature, 24°C ± 4°C.
4. Dump contents of wells into waste. Blot and gently but firmly tap over absorbent paper to remove most of the residual liquid. Overly aggressive banging of the
plate or use of vacuum aspiration devices in an attempt to remove all residual liquid is not necessary and may cause variable dissociation of antibody bound material resulting in lower ODs and worse precision. Fill wells generously to overflowing with diluted wash solution using a squirt bottle or by pipetting in ~350µL. Dump and tap again. Repeat for a total of 4 washes. Wipe off any liquid from the bottom outside of the microtiter wells as any residue can interfere in the reading step.
Do not allow wash solution to remain in wells for longer than a few seconds. Do not allow wells to dry before adding substrate.
5. Pipette 100uL of TMB substrate.
6. Incubate at room temperature for 30 minutes. DO NOT SHAKE.
7. Pipette 100uL of Stop Solution.
8. Read absorbance at 450/650nm.