Purified proteins or peptides are ideal as controls in flow cytometry, Western blot, and ELISA. Human Foreskin Fibroblast Whole Cell Lysate Positive Control for Anti-Epitope Tag Antibodies Recommended Control Cell Lysates and Nuclear Extracts: EGF Stimulation Controls All extracts are tested by SDS-PAGE using 4–20% gradient gels and immunoblot analysis using antibodies to key cell signaling components to confirm the presence of both high molecular weight and low molecular weight proteins.
Additionally, Rockland offers a variety of lysates that contain over-expressed proteins (tagged and untagged) that can serve as positive controls for antibody reactivity. Lysates are also available from normal animal tissue derived from primary organs such as liver, heart, and brain. Control lysates may be from cells that are stimulated with insulin, doxorubicin, etoposide, nocodozole, TNFa, or EGF. Lysates are generated from either whole cells, which contain cell membrane, cytoplasmic, and nuclear proteins, or nuclear extracts, which are predominantly proteins that originate in the nucleus. Our ready-to-use whole-cell lysates and nuclear extracts are derived from cell lines or tissues using highly advanced extraction protocols to ensure high quality, protein integrity, and lot-to-lot reproducibility. Rockland offers control cell lysates and nuclear extracts for use on SDS-PAGE as standalone samples or in combination with antibodies in Western blotting experiments. Recommended Loading Control Antibodies: Western Blot (WB)Į.coli Combined HCP Biotin Conjugated AntibodyĬontrol Cell Lysates and Nuclear Extracts Rockland’s loading control antibodies are suitable in assays including ELISA, FLISA, Western blot, IF, and IHC.
POSITIVE CONTROL NEGATIVE CONTROL VERIFICATION
Loading control antibodies not only allow the verification of equal protein loading between samples in Western blot assays, but they also allow for identification of certain cell compartmentalization or cellular localization in immunofluorescence microscopy (IF) and immunohistochemistry (IHC). In Western blot assays, the loading control should be at a different molecular weight than the protein of interest, as this allows the protein to be visually distinguishable. Loading controls are essential for the interpretation of assays.
Ideal loading controls are expressed constitutively and at high levels with low variability between cell lines and experimental conditions. Loading control antibodies mostly recognize housekeeping proteins in cells used in a scientific experiment and allow the verification of equal protein loading between samples.
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