Home  ■  Participation  ■  Assessments  ■  Epitopes  ■  Protocols  ■  Techniques  ■  Links
Tissue handling ■  Section handling  ■  Epitope retrieval  ■  Endogenous biotin
Primary antibody  ■  Visualization  ■  Chromogene  ■  Controls  ■  Recommended control tissue

Epitope retrieval CD117

The influence of HIER in the detection of CD117 according to the experience at Institute of Pathology, Aalborg Hospital, is illustrated.

For DakoCytomation's polyclonal antibody (Ab) A 4502, we recommend a dilution of 1:100 after HIER in Tris/EDTA pH9. Even an Ab dilution of 1:500 gives in all cases a stronger staining after HIER than an Ab dilution of 1:100 without pretreatment does (also see Assessments).

The recommended protocol may give a slight overstaining in tumours with many epitopes (Fig. 1) as well a very faint background staining. However, for the staining of tumours with few epitopes (Fig. 2) the method is superior.
Fig. 1A. GIST 1 stained for CD117. Pretreatment: HIER in Tris/EDTA pH 9. Ab diluted 1:100  The staining of perivascular cells is intense while peripheral cells show a weaker staining. Fig. 1B. GIST 1 stained for CD117. Pretreatment: HIER in Tris/EDTA pH 9. Ab diluted 1:500. The staining is still acceptable. Fig. 1C. GIST 1 stained for CD117. No pretreatment. Ab diluted 1:100. The staining is weaker than in Fig. 1 B, particularly in the peripheral cells.
Fig. 2A. GIST 2 stained for CD117. Pretreatment: HIER in Tris/EDTA pH 9. Ab diluted 1:100. Moderate staing of scattered tumour cells (in other areas of the tumour, a staining intensity similar to that of Fig. 1 was revealed). Fig. 2B. GIST 2 stained for CD117. Pretreatment: HIER in Tris/EDTA pH 9. Ab diluted 1:500. The staining is barely perceptible. Fig. 2C. GIST 2 stained for CD117. No pretreatment: Ab diluted 1:100. The staining is lost.
Fig. 3A. Normal skin stained for CD117. Pretreatment: HIER in Tris/EDTA pH 9. Ab diluted 1:100. Strong staining of mastcells and melanocytes. Fig. 3B. Normal skin stained for CD117. Pretreatment: HIER in Tris/EDTA pH 9. Ab diluted 1:500. The staining of mastcells and melanocytes is acceptable. Fig. 3C. Normal skin stained for CD117. No pretreatment. Ab diluted 1:100. The mastcells and melanocytes.show a slightly weaker staining than in Fig. 3B.
MV/SN/AS

Last update 21-09-2005