MLK4 alpha in Human Colon Cancer Tissue.
MLK4 alpha was detected in immersion fixed paraffin-embedded sections of human colon cancer tissue using 10 µg/mL Human/Mouse/Rat MLK4 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3435) overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained with the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). Specific labeling was localized to microvilli of epithelial cells. Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human/Mouse/Rat MLK4 alpha by Western Blot.
Western blot shows lysates of K562 human chronic myelogenous leukemia cell line, Ramos human Burkitt's lymphoma cell line, A20 mouse B cell lymphoma cell line, CH‑1 mouse B cell lymphoma cell line, Rat‑2 rat embryonic fibroblast cell line, and L6 rat myoblast cell line. PVDF membrane was probed with 1 µg/mL of Human/Mouse/Rat MLK4 alpha Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3435) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for MLK4 alpha at approximately 62 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: MLK4 alpha
Mixed-lineage kinases (MLKs) function as mitogen-activated protein kinase kinase kinases (MAP3Ks), activating the p38 and JNK signaling cascades. The potential importance of MLK4’s role in cell regulation has been indicated by the finding of nine separate mlk4 point mutations in a recent sampling of colorectal cancers. MLK4 has two splice variants, with MLK4 alpha containing a different and shorter C-terminus than MLK4 beta.
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