Immunohistochemical analysis of paraffin-embedded human urothelial carcinoma using Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb.
Chromatin immunoprecipitations were performed with cross-linked chromatin from NCI-H2052 cells and either TAZ (E8E9G) Rabbit mAb #83669 or Normal Rabbit IgG #2729 using SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads) #9005. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human CTGF Promoter Primers #14927, human SMYD3 intron 2 primers, and SimpleChIP® Human CTGF Upstream Primers #14928. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Immunohistochemical analysis of paraffin-embedded SK-MEL-28 cell pellet (left, positive) or RL cell pellet (right, negative) using Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb.
Confocal immunofluorescent analysis of MCF 10A cells (high-expressing) at low (left) and high-confluence (middle) versus SH-SY5Y cells (right, low-expressing) using TAZ (E8E9G) Rabbit mAb (green). Nuclei are labeled with DAPI #4083 (blue). Note that decreased nuclear localization of TAZ protein is seen in high-confluence MCF 10A cells.
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma, untreated (left) or lambda phosphatase treated (right), using Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb.
Immunoprecipitation of TAZ protein from HeLa cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is TAZ (E8E9G) Rabbit mAb. Western blot analysis was performed using TAZ (D3I6D) Rabbit mAb #70148. Mouse anti-rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used for detection to avoid cross-reactivity with IgG.
Immunohistochemical analysis of paraffin-embedded human lung adenocarcinoma using Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb.
Western blot analysis of extracts from various cell lines using TAZ (E8E9G) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Expression levels of TAZ among cell lines are consistent with expectations based on publicly available bioinformatic databases.
Immunohistochemical analysis of paraffin-embedded mouse colon using Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse lung using Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human ovarian clear cell carcinoma using Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb.
Immunoprecipitation of Phospho-TAZ (Ser89) from A-204 cell extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb. Western blot analysis was performed using Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb. Mouse Anti-Rabbit IgG (Conformation Specific) (L27A9) mAb (HRP Conjugate) #5127 was used as the secondary antibody. Asterisk (*) indicates weak detection of phosphorylated YAP, due to sequence similarity in the regions surrouding TAZ (Ser89) and YAP (Ser127).
Western blot analysis of extracts from PANC-1 and A-204 cells, untreated (-) or treated with calf intestinal phosphatase (CIP) and λ phosphatase (+) using Phospho-TAZ (Ser89) (E1X9C) Rabbit mAb (upper), TAZ (D3I6D) Rabbit MAb #70148 (middle) and β-Actin (D6A8) Rabbit mAb #8457 (lower). Asterisk (*) indicates weak detection of phosphorylated YAP, due to sequence similarity in the regions surrounding TAZ (Ser89) and YAP (Ser127).
PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.
TAZ is a transcriptional co-activator with a PDZ-binding motif that is regulated by its interaction with 14-3-3 proteins (1). TAZ shares homology with the WW domain of Yes-associated protein (YAP) (1). TAZ is proposed to modulate the switch between proliferation and differentiation of mesenchymal stem cells (MSC) via interaction with transcription factors Runx2 and PPARγ. This process is critical to normal tissue development and the prevention of tumor formation. Due to its role in determination of MSC fate, TAZ may have clinical relevance to several human diseases caused by an imbalance of MSC differentiation (2,3). TAZ is negatively regulated via phosphorylation by LATS1/2, core kinases in the Hippo signaling pathway that controls stem cell development, tissue growth and tumor development (4).
Phosphorylation of TAZ at Ser89 functions to destabilize TAZ protein by promoting 14-3-3 binding, cytoplasmic sequestration, and proteasomal degradation, thereby reducing the ability of TAZ to co-activate transcription of downstream target genes. Mutation of Ser89 to alanine (S89A) yields a constitutively active form of TAZ; expression of TAZ (S89A) in breast cancer cells was shown to promote a cancer stem cell phenotype (5).
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