Advanced Solutions
for Advanced Pathology
Phosphohistone H3 (PHH3) (Polyclonal)
Rabbit Polyclonal Antibody
Cat. No. Description
Volume
45324 IMPATH Phosphohistone H3 RTU R (PHH3)
50 Tests
44370 Phosphohistone H3 RTU R (PHH3)
7 ml Ready To Use
44759 Phosphohistone H3 0,1 R (PHH3)
100 µl liquid Concentrated
44760 Phosphohistone H3 1 R (PHH3)
1 ml liquid Concentrated
Product Specifications
Designation
IVD
Reactivity
Paraffin
Visualization
Nuclear
Control
Tonsil
Stability
Up to 36 mo. at 2-8°C
Manual Protocol*
• Pretreatment: Heat Induced Epitope
Retrieval (HIER)
• Primary Antibody Incubation Time:
10-30min @ 25-37°C
• 2-step polymer detection
*Please refer to product insert for complete protocol.
ImPath Protocol*
• Dewax: Dewax Solution 2 (DS2)
• Pretreatment: Retrieval Solution pH 9.0
(TR1) 32min @ 98-103°C
• Primary Antibody Incubation Time:
10-90min @ 25-37°C
• HRP Polymer (Universal) or AP Polymer
(Universal) for 12 min
*Please refer to product insert for complete protocol.
Product Description
Phosphohistone H3 (PHH3) is a core histone protein, which together with other histones, forms the major protein constituents of the chromatin
in eukaryotic cells. In mammalian cells, phosphohistone H3 is negligible during interphase but reaches a maximum for chromatin condensation
during mitosis. Immunohistochemical studies showed anti-PHH3 specifically detected the core protein histone H3 only when phosphorylated at
serine 10 or serine 28. Studies have also revealed no phosphorylation on the histone H3 during apoptosis. Therefore, anti-PHH3 can serve as a
mitotic marker to separate mitotic figures from apoptotic bodies and karyorrhectic debris, which may be a very useful tool in diagnosis of tumor
grading and staging, especially in central nervous system tumors, melanomas, soft tissue sarcomas, and gastrointestinal stromal tumor. PHH3
index is a sensitive measure of mitotic activity and has several technical advantages over the currently existing methods.
The PHH3 mitotic index method provides a true index, is independent of tumor cellularity (which can confound the scoring of mitoses per unit
area), and greatly facilitates the identification of mitotic figures compared to the traditional H&E staining method. Because it stains only cells in
mitosis, PHH3 offers the possibility of obtaining a true mitotic index, compared to Ki-67 proliferation index, which is positively stained in cells
in all phases of the cell cycle, except G0. This feature of PHH3 staining also makes it less susceptible to variation based on antibody dilution or
labeling intensity, as the pathologist can confirm that PHH3-labeled cells are also mitotic based on nuclear morphology.
Comparison of immunoreactivity of PHH3 and Ki-67 in cell cycle
Cell Cycle
PHH3
Ki-67
G0 phase
-
-
Interphase
G1 phase
-
+
S phase
-
+
G2 phase
-
+
Mitosis phase
Prophase
+
+
Metaphase
+
+
Anaphase
+
+
Telophase
+
+
Reference
1. Gurley LR, et al. Eur J Biochem. 1978; 84:1-15.
2. Hendzel MJ, et al. J Biol Chem. 1998; 273:24470-8.
3. Colman H, et al. Am J Surg Pathol. 2006; 30:657-64.
4. Nasr MR, El-Zammar 0. Am J Dermatopathol. 2008; 30:117-22.
5. Kim YJ, et al. Am J Clin Pathol. 2007; 128:118-25.
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