Trs high ph

Manufactured by Agilent Technologies

The TRS High pH is a laboratory equipment product manufactured by Agilent Technologies. It is designed to perform high-pH analysis and separation tasks. The core function of this product is to enable efficient and accurate measurement and analysis of samples with high pH levels.

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Lab products found in correlation

Envision hrp detection system, by Agilent Technologies (2 mentions) Bcl 2 oncoprotein, by Agilent Technologies (1 mentions) Dako envision flex high ph detection kit, by Agilent Technologies (1 mentions) Autostainer plus, by Agilent Technologies (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions)

3 protocols using trs high ph

Immunohistochemical Staining for Bmi-1, CK15, and Bcl-2

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Immunohistochemical staining was carried out according to a standard method. 3-μm tissue sections were deparaffinised in xylene and rehydrated through a graded alcohol series. Heating in a microwave oven in a target retrieval solution pH 9.0 (TRS High pH; Dako) for 30 min was used for antigen retrieval. Endogenous peroxidase activity was quenched with 0.3% hydrogen peroxide in methanol for 30 min. The sections were washed with TBS and incubated with primary antibodies against: Bmi-1 (Invitrogen, USA, dilution 1 : 600, Catalog number PA5-23308), and CK15 (Invitrogen, USA, dilution 1 : 400, Catalog number MA1-90926), Bcl-2 Oncoprotein (Dako, RTU- Flex, clone 124, Catalog number IR 614). After washing, an adequate EnVision-HRP detection system (Dako, Carpinteria, CA, USA) was used. 3,3’-diaminobenzidine was used as the chromogen. After counterstaining with Mayer’s haematoxylin, the slides were washed, dehydrated, cleared in xylene and coverslipped. Negative controls for immunohistochemical staining were prepared with primary antibodies replaced by the antibody diluent.

Iljin A., Stasikowska-Kanicka O., Zieliñski T., Bąkiewicz A., Sporny S., Woźniak-Roszkowska E, & Antoszewski B. (2022). Immunoexpression of Bmi-1, CK15, Bcl-2 in different types of basal cell carcinomas. Advances in Dermatology and Allergology/Postȩpy Dermatologii i Alergologii, 39(5), 980-985.

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Immunohistochemical Staining of SOX2 and TAZ

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Immunohistochemical staining was carried out according to a standard method. 3-µm tissue sections were deparaffinized in xylene and rehydrated through a graded alcohol series. Heating in a microwave oven in a solution of target retrieval solution pH 9.0 (TRS High pH; Dako), for 30 minutes was used for antigen retrieval. Endogenous peroxidase activity was quenched with 0,3% hydrogen peroxide in methanol for 30 minutes. The sections were washed with TBS and incubated all night with polyclonal rabbit primary antibodies against: SOX2 (ThermoFisher Scientific, USA, dilution 1:300, Catalog number PA1-094), and TAZ (Abcam, UK, dilution 1:400, Catalog number ab84927). The sections for α-SMA staining were incubated 30 minutes with monoclonal mouse primary antibodies against actin (Dako; clone: 1A4, RTU) After washing, an adequate EnVision-HRP detection system (Dako, Carpinteria, CA, USA) was used. 3,3'-diaminobenzidine was used as the chromogen. After counterstaining with Mayer's hematoxylin, the slides were washed, dehydrated, cleared in xylene and coverslipped. The negative controls for immunohistochemical staining were prepared with the primary antibodies replaced by the antibody diluent.

Stasikowska-Kanicka O., Woskowicz M., Ulewicz K., Danilewicz M, & Wągrowska-Danilewicz M. (2021). Close relationship between TAZhigh/SOX2high co-localization and metastasis in oral squamous cell carcinoma. Polish journal of pathology : official journal of the Polish Society of Pathologists, 72(3).

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Immunohistochemical Staining Protocol for CLPTM1L

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IHC staining was performed on the Dako Autostainer Plus using the Dako EnVision^™ FLEX High pH Detection Kit (K8010) (Dako, Carpinteria, CA). Slides were deparaffinized to DI water. Antigen Retrieval was performed on Dako PT Link water bath. The antigen retrieval was done at 97°C for 20 minutes. The slides were cooled until they reached 65°C. All slides for all antibodies were placed in Tris/EDTA pH 9 (Dako TRS High pH). Slides were washed in Dako wash buffer for 5 minutes. Slides were subjected to a peroxidase Block for 5 minutes. Slides were rinsed twice with wash buffer. Slides were incubated with primary antibody CLPTM1L (rabbit polyclonal, Sigma Aldrich cat# HPA014791, lot A57952) diluted to 1:400 for 30 minutes. Slides were rinsed with wash buffer. Slides were incubated with secondary antibody for 20 minutes and rinsed twice with wash buffer. Slides were incubated with DAB substrate for 10 minutes and rinsed with wash buffer. Slides were stained with hematoxylin for 7 minutes and rinsed with DI water. Slides were dehydrated and coverslipped for viewing. Staining intensity in tumor tissue was scored by three pathologists and averaged. Tumors were scored as negative – 0, weak – 1, intermediate – 2, or strong – 3. Independent scores were averaged. Antibodies for IHC on xenograft tumors were Ki-67 (CRM 325 A,B,C) and Cleaved Caspase-3 (Asp175, Cell Signaling, Boston MA) 1:300.

Puskás L.G., Mán I., Szebeni G., Tiszlavicz L., Tsai S, & James M.A. (2016). Novel Anti-CRR9/CLPTM1L Antibodies with Antitumorigenic Activity Inhibit Cell Surface Accumulation, PI3K Interaction, and Survival Signaling. Molecular cancer therapeutics, 15(5), 985-997.

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