IHC for cleaved PARP (cPARP) was performed using a rabbit monoclonal antibody specific for PARP cleaved at Asp214. Tissue sections were deparafinised and rehydrated prior to antigen retrieval using High pH (pH9.0) Target Antigen Retrieval Solution (K8004, Dako UK Ltd), in PT-LINK (PT101, Dako UK Ltd). Tissue sections were stained for cPARP using rabbit monoclonal anti-cPARP antibody (Asp214, clone D64E10, Cell Signaling Technology, #5625) diluted 1:100 in EnVision Antibody Diluent (K8006, Dako UK Ltd). Sections were washed using wash buffer (Dako,S3006) before colour development using anti-rabbit link reagent EnVision FLEX LINKER (K8019, Dako UK Ltd). Tissues sections were counter stained with hematoxylin and coverslips mounted using DPX. The percentage of cPARP positive cells was calculated from a minimum of 500 invasive tumour cells. If fewer than 500 invasive tumour cells were present the sample was recorded as insufficient invasive tumour (IIT).
Envision flex linker
Manufactured by Agilent Technologies
Sourced in United Kingdom
The EnVision FLEX LINKER is a laboratory equipment designed to connect multiple instruments within the EnVision FLEX platform. Its core function is to enable seamless integration and communication between compatible Agilent instruments, facilitating efficient workflow and data management.
Automatically generated - may contain errors
Lab products found in correlation
Pt link, by Agilent Technologies (2 mentions)
Wash buffer, by Agilent Technologies (2 mentions)
Envision antibody diluent, by Agilent Technologies (2 mentions)
3 3 diaminobenzidine (dab), by Agilent Technologies (1 mentions)
Envision flex peroxidase blocking reagent, by Agilent Technologies (1 mentions)
Bcl2 clone 124, by Agilent Technologies (1 mentions)
Pt link pre treatment system, by Agilent Technologies (1 mentions)
3 protocols using envision flex linker
1
Immunohistochemistry for Cleaved PARP in Tumours
2
Immunohistochemistry for Cleaved PARP in Tumours
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IHC for cleaved PARP (cPARP) was performed using a rabbit monoclonal antibody specific for PARP cleaved at Asp214. Tissue sections were deparafinised and rehydrated prior to antigen retrieval using High pH (pH9.0) Target Antigen Retrieval Solution (K8004, Dako UK Ltd), in PT-LINK (PT101, Dako UK Ltd). Tissue sections were stained for cPARP using rabbit monoclonal anti-cPARP antibody (Asp214, clone D64E10, Cell Signaling Technology, #5625) diluted 1:100 in EnVision Antibody Diluent (K8006, Dako UK Ltd). Sections were washed using wash buffer (Dako,S3006) before colour development using anti-rabbit link reagent EnVision FLEX LINKER (K8019, Dako UK Ltd). Tissues sections were counter stained with hematoxylin and coverslips mounted using DPX. The percentage of cPARP positive cells was calculated from a minimum of 500 invasive tumour cells. If fewer than 500 invasive tumour cells were present the sample was recorded as insufficient invasive tumour (IIT).
3
Immunohistochemical Analysis of BCL2 in Murine Lymphoma
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Sections from formalin-fixed, paraffin-embedded blocks from murine lymphoma xenografts were cut and stained with hematoxylin and eosin. IHC was performed using BCL2 primary antibody (clone 124, Mouse, Dako). Deparaffinization, rehydration, and antigen retrieval was performed with EnVision FLEX Target Retrieval (1:50, High pH) in the PT link pretreatment system (Dako). After blocking of endogenous peroxidase activity (EnVision FLEX Peroxidase blocking reagent, Dako S2001), the samples were incubated with primary antibody BCL2 (clone 124, 1:100, Dako) for 50 minutes at room temperature. Amplification of signal of primary antibody was performed with EnVision FLEX+ Linker and the reaction was visualized by chromogen substrate DAB (3,30-diaminobenzidine tetrahydrochloride; Dako). Nuclei were counterstained with Harris hematoxylin. After dehydration, slides were mounted in organic solvent–based medium and evaluated under light microscope. The slides were evaluated by expert hematopathologist. Samples were considered positive if ≥ 30% of the tumor cells showed cytoplasmic positivity.
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