Leica HMB45 Manuel D’Utilisation

PA0027 
Page 3

•  Consult Federal, State or local regulations for disposal of any potentially toxic components.
•  Minimize microbial contamination of reagents or an increase in non-specific staining may occur.
•  Retrieval, incubation times or temperatures other than those specified may give erroneous results. Any such change must be 

validated by the user.

Melanoma Marker (HMB45) primary antibody is recommended for use on an automated Bond system in combination with either 

Bond Polymer Refine Detection (DS9800) or Bond Polymer Refine Red Detection (DS9390). The recommended staining protocols for 

Melanoma Marker (HMB45) primary antibody are IHC Protocol F when using Bond Polymer Refine Detection and IHC Protocol J when 

using Bond Polymer Refine Red Detection. Enzyme pretreatment is recommended using Bond Enzyme 1 for 5 minutes.

Normal Tissues
Melanoma Marker (HMB45) showed positive granular, cytoplasmic staining in melanocytes in the basal layer of epidermis. All other 

normal tissues were negative. (Total number of cases stained = 99).
Tumor Tissues 
Melanoma Marker (HMB45) stained 13/13 malignant melanomas. No staining was observed in a variety of other tumors. (Total number 

of cases stained = 101).
Melanoma Marker (HMB45) is recommended for use as part of a panel of antibodies for the diagnosis of melanomas.

Melanoma Marker (HMB45) is recommended for use with either Bond Polymer Refine Detection or Bond Polymer Refine Red Detection 

and Bond ancillary reagents. Users who deviate from recommended test procedures must accept responsibility for interpretation 

of patient results under these circumstances. The protocol times may vary, due to variation in tissue fixation and the effectiveness 

of antigen enhancement, and must be determined empirically. Negative reagent controls should be used when optimizing retrieval 

conditions and protocol times.

Refer to reference 3 for remedial action.
Contact your local distributor or the regional office of Leica Biosystems to report unusual staining.

Further information on immunostaining with Bond reagents, under the headings Principle of the Procedure, Materials Required, 

Specimen Preparation, Quality Control, Assay Verification, Interpretation of Staining, Key to Symbols on Labels, and General Limitations 

can be found in “Using Bond Reagents” in your Bond user documentation.

1.  Clinical Laboratory Improvement Amendments of 1988, Final Rule 57 FR 7163 February 28, 1992.
2.  Villanova PA. National Committee for Clinical Laboratory Standards (NCCLS). Protection of laboratory workers from infectious 

diseases transmitted by blood and tissue; proposed guideline. 1991; 7(9). Order code M29-P.

3.  Bancroft JD and Stevens A. Theory and Practice of Histological Techniques. 4th Edition. Churchill Livingstone, New York. 1996.
4.  Kapur RP, Bigler SA, Skelly M, et al. Anti-melanoma monoclonal antibody HMB45 identifies an oncofetal glycoconjugate associated 

with immature Melanoma Markers. The Journal of Histochemistry and Cytochemistry. 1992; 40(2):207-212.

5.  Gown AM, Vogel AM, Hoak D, et al. Monoclonal antibodies specific for melanocytic tumours distinguish subpopulations of 

melanocytes. American Journal of Pathology. 1986;123(2):195-203.

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09 May 2013