Name: FFPE antigen retrieval and staining kit
SKU: BCASFP
Price: 195.00 USD
Availability: InStock

Formaldehyde crosslinks are formed between protein molecules, in particular with the basic amino acid – lysine. Only those lysine residues on the exterior of the protein molecule can react with formaldehyde. Formaldehyde crosslinks are made primarily on the hydrophilic areas of proteins. The reaction between aldehyde and protein is pH-dependent, progressing more rapidly at higher pH values.

Formaldehyde concentrations ranging from 4-10% are often used in fixation buffer. BiCell Scientific has found that lower formaldehyde concentrations such as 1-2% can better preserve antigenicity while higher formaldehyde concentrations such as 4-10% retain good tissue morphology. Formalin-fixed paraffin-embedded (FFPE) sections (formalin: 10% formaldehyde), however, require antigen retrieval processes to revive antigenicity in tissues, which is essential to IHC applications.

The kit contains:

250ml of Universal antigen retrieval buffer (proprietary)
50ml of FFPE autofluorescence quenching buffer (proprietary)
100ml of Block buffer (bovine serum, Tween-20, NaCl and Tris buffered saline)
500ml of Wash buffer (Tween-20, NaCl, and Tris buffered saline)
1ml of Mounting buffer (Mowiol and Tris buffered saline)

Store at -20^oC

For Research Use Only. Not for use in clinical diagnostics.

Thaw kit content to room temperature.
Deparaffinize FFPE sections with Xylene.
Rehydrate sections with decreasing concentrations of ethyl alcohol (100% to 70%).
Wash slides 2 times with dH₂O.
Dip slides into universal antigen retrieval buffer.
Microwave sections at boiling temperature for 10min.
Wash slides 2 times with dH₂O.
Dip slides into FFPE autofluorescence quenching buffer for 20min
Wash slides with 70% ethyl alcohol
Wash slides with 1xPBS
Drain Wash solution against a towel and put slides into the wet chamber.
Dilute primary antibody at 1:50 in Block solution and add to each slide (300-500 µl is sufficient to cover the section on each slide).
Incubate the slides in the wet chamber at room temperature for 2hr.
Take slides out of the wet chamber, drain off antibody, and wash three times with the Wash solution.
Dilute secondary antibodies at 1: 200 in Block solution and add to each slide and put slides back into the wet chamber.
Incubate the slides in the wet chamber at room temperature for 1hr.
Take slides out of the wet chamber, drain off antibody, and wash three times with the Wash solution.
If using fluorophore labeled secondary antibodies, then add 20 ul of Mowiol solution to the section on each slide. Place a cover glass onto the section and apply a small pressure to remove any trapped air bubble. Let Mowiol solution dry at room temperature for 1hr. Store glass slides at 4^oC for less than 1 month.

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FFPE antigen retrieval and staining kit

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