Immunohistochemistry (IHC) was performed on representative whole formalin fixed, paraffin embedded (FFPE) 5 micron-thick tissue sections. Heat induced epitope retrieval was performed with FLEX TRS Low pH Retrieval buffer (6.10; Dako, Carpinteria, CA) for 20 minutes. After peroxidase and protein blocking, the rabbit polyclonal FOXI1 antibody (1:250; Atlas Antibodies, Stockholm, Sweden) or rabbit polyclonal RHCG antibody (1:4000; LSBio, Seattle, WA) was applied to the sections and incubated at room temperature for 60 minutes. The FLEX + Rabbit EnVision System (Dako, Carpinteria, CA) and DAB chromogen were used for detection. Immunopositivity was defined as brown pigmentation in the membrane/cytoplasm (for RHCG) or nucleus (for FOXI1) of tumor cells. The staining was independently assessed by two pathologists (SLS and RM) in each case for presence and pattern of expression.
Flex trs low ph retrieval buffer
Manufactured by Agilent Technologies
The FLEX TRS Low pH Retrieval buffer is a laboratory reagent used to prepare samples for immunohistochemical (IHC) analysis. It is designed to facilitate the retrieval of antigenic epitopes that may have been masked or altered during the fixation process. The buffer operates at a low pH to help unmask these epitopes, allowing for improved detection and visualization of target proteins in tissue samples.
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Lab products found in correlation
2 protocols using flex trs low ph retrieval buffer
1
Immunohistochemical Analysis of FOXI1 and RHCG
2
Immunohistochemistry for CXCR4 and CD99 Evaluation
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Immunohistochemistry for CXCR4 and CD99 was performed on formalin fixed paraffin embedded tumor sections using the Dako Autostainer Link (Dako, Carpinteria, CA). Following rehydration, sections were treated with heat induced epitope retrieval (HIER). For CXCR4 HIER was performed with FLEX TRS Low pH Retrieval buffer (pH 6.10) (Dako) for 20 minutes. For CD99 HIER was performed with FLEX TRS High pH Retrieval buffer (pH 9.01) (Dako) for 20 minutes. After peroxidase blocking, CXCR4 antibody (ab124824; abcam, Cambridge, MA) was applied at a dilution of 1:4000 at room temperature for 60 minutes, and CD99 antibody (M3601; Dako, Carpinteria, CA) was applied at a dilution of 1:100 at room temperature for 60 minutes. The FLEX HRP EnVision System (Dako) was used for detection. DAB chromagen was then applied for 10 minutes. Slides were counterstained with Harris Hematoxylin for 5 seconds and then dehydrated and coverslipped.
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