Western blot analysis of total cell lysates and conditioned medium from L cells overexpressing mouse Wnt3a, compared with parental L cells, using Wnt3a (C64F2) Rabbit mAb.
Western blot analysis of total cell lysates from HeLa, LN18, PANC1 cells and HeLa cells conditioned medium (CM) using Wnt5a/b (C27E8) Rabbit mAb.
Western blot analysis of total cell lysates from HeLa cells untreated, treated 5 hours with conditioned medium (CM) from L cells, or treated 5 hours with conditioned medium from L cells overexpressing Wnt3a, using Phospho-LRP6 (Ser1490) Antibody #2568.
Western blot analysis of total cell lysates from various cell lines using Dvl2 (30D2) Rabbit mAb.
Western blot analysis of extracts from Hep G2 and HeLa cells using LRP6 (C47E12) Rabbit mAb.
Western blot analysis of extracts from NIH/3T3, C6, A549, BAEC and COS cells using Dvl3 Antibody.
Western blot analysis of total cell lysates from HepG2 and SW620 cells using Naked1 (C30F10) Rabbit mAb.
Western blot analysis of extracts from various cell lines using Naked2 (C67C4) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Axin1 (C76H11) Rabbit mAb.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of total cell lysates from Rat2 and NIH/3T3 cells, untreated or treated with conditioned medium containing Wnt3a for 60 minutes and also treated with or without calf intestinal phosphatase (CIP), using Dvl2 (30D2) Rabbit mAb.
Western blot analysis of extracts from various cell lines using Axin1 (C76H11) Rabbit mAb.
|Wnt3a (C64F2) Rabbit mAb 2721||20 µl||
|Wnt5a/b (C27E8) Rabbit mAb 2530||20 µl||
|Phospho-LRP6 (Ser1490) Antibody 2568||20 µl||
|Dvl2 (30D2) Rabbit mAb 3224||20 µl||
||H M R Mk||90 to 95||Rabbit|
|LRP6 (C47E12) Rabbit mAb 3395||20 µl||
||H M||180, 210||Rabbit IgG|
|Dvl3 Antibody 3218||20 µl||
||H M R Hm Mk Mi B||88 to 93||Rabbit|
|Naked1 (C30F10) Rabbit mAb 2201||20 µl||
||H||59, 61||Rabbit IgG|
|Naked2 (C67C4) Rabbit mAb 2073||20 µl||
|Axin1 (C76H11) Rabbit mAb 2087||20 µl||
||H M||110||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
The Wnt Signaling Antibody Sampler Kit provides an economical means of detecting integral proteins within the Wnt signaling pathway. The kit contains enough primary and secondary antibody to perform two Western blots with each.
Wnt3a (C64F2) Rabbit mAb detects transfected Wnt3a protein in L cells and the conditioned medium from L/Wnt3a cells. Phospho-LRP6 (Ser1490) Antibody detects endogenous levels of LRP6 protein when phosphorylated at Ser1490. The remaining antibodies in the Wnt Signaling Sampler Kit detect endogenous levels of their respective total proteins.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu256 of human Wnt3a, Glu176 of human Wnt5a, Met988 of human LRP6, Gly176 of human Dvl2, Leu53 of human Naked1, and Ala217 of human Naked2. The monoclonal antibody for Axin1 is produced by immunizing animals with recombinant human Axin1. The phospho-specific polyclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser1490 of human LRP6. The total polyclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding the carboxy terminus of human Dvl3. Polyclonal antibodies are purified by Protein A and peptide affinity chromotagraphy.
The Wnt family includes several secreted glycoproteins that play important roles in animal development (1). Aberrant activation of Wnt signaling pathways is involved in several types of cancers (2). Wnt members bind to the Frizzled family of proteins and activate several signaling pathways (3). The canonical Wnt/β-catenin pathway requires a coreceptor from proteins belonging to the low-density lipoprotein receptor (LDLR) family, such as LRP5 and LRP6 (4). Upon stimulation with Wnt, LRP6 is phosphorylated by kinases such as GSK-3 and CK1 and subsequently recruits Axin to the membrane (5-7). In the absence of Wnt stimulation, the multidomain scaffold proteins Axin1 and Axin2 negatively regulate Wnt signaling. Axin1 complexes with APC, GSK-3β, Dvl and β-catenin and promotes the GSK-3β-mediated phosphorylation and subsequent degradation of β-catenin (8-9). The Disheveled (Dsh) proteins (Dvl1, Dvl2 and Dvl3) inhibit glycogen synthase kinase-3β, thereby promoting β-catenin stabilization (4,10). Naked inhibits the canonical Wnt/β-catenin pathway by binding to Dsh proteins and directing Dsh activity towards the planar cell polarity pathway (11-12).
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