Generation of cell type-specific monoclonal antibodies for the planarian and optimization of sample processing for immunolabeling

Table 2 Optimization of whole animal sample processing

mAb	Tissue specificity	Fixation protocol
					Mg	Mg	Mg
		HCl	HCl	NAc	HCl	HCl	NAc
		Mcn	FA	FA	Mcn	FA	FA
2G4	Intestine & epidermis	+	+	+	++	+	+
3F11	None (MeOH bleach)	-	-	-	-	-	-
3F11	Intestine (PBS bleach)	-	-	-	-	+	+
3G9	Intestine	-	+	+	++	+	+
4D2	Intestine	+	-	-	++	+	+
1E12	Nuclei, elevated in epidermis and intestine	+	+	-	+	-	-
1H8	Nuclei, elevated in CNS	-	-	+	-	+	+
2C11	Peripharyngeal secretory cells and processes	-	+	+	-	+	++
2D2	Subset of neurons and processes	-	+	++	-	+	++
2H3	Muscles	++	+	++	++	+	++
3G7	Epidermal basement membrane	-	+	-	-	+	-
3H3	Epidermis, enriched in cell:cell junctions	++	+	+	++	-	-

Qualitative assessment of signal:noise for the predominant tissue labeled by each mAb. “-“ indicates negligible or non-uniform labeling, poor morphology, and/or high background. “+“ indicates moderate specificity, uniform labeling, and/or robust labeling accompanied by moderate background. “++” indicates robust signal with minimal noise.

ISSN: 1471-213X