Human/Mouse/Rat MLK4 alpha Antibody

Catalog #: AF3435 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF3435
AF3435-SP

MLK4 alpha in Human Colon Cancer Tissue.

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Human/Mouse/Rat MLK4 alpha Antibody Summary

Species Reactivity

Human, Mouse, Rat

Specificity

Detects endogenous human, mouse, and rat MLK4 alpha. Because the peptide immunogen corresponds to a region at the C-terminus of MLK4 alpha absent in MLK4 beta, no reactivity with MLK4 beta is present.

Source

Polyclonal Rabbit IgG

Purification

Antigen Affinity-purified

Immunogen

Human MLK4 alpha synthetic peptide (aa 560 - 570)
ELSALPPGLLC
Accession # CAC84639

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunohistochemistry

5-15 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunohistochemistry

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MLK4 alpha in Human Colon Cancer Tissue. MLK4a was detected in immersion fixed paraffin-embedded sections of human colon cancer tissue using 10 µg/mL Human/Mouse/Rat MLK4a 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.

Western Blot

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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 MLK4a Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3435) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for MLK4a at approximately 62 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Long Name

Mixed Lineage Kinase 4 alpha

Entrez Gene IDs

84451 (Human)

Alternate Names

dJ862P8.3; KIAA1804; MAP3K21; MLK4 alpha; MLK4