IHC, HMB45 Test

HMB stands for Human Melanoma Black and HMB-45 is a monoclonal antibody which reacts against the antigen that is present in melanocytic tumors like melanomas. It is used in the pathologies as a marker for the detection of the tumors associated with melanoma. Pmel 17 is the specific antigen that recognized by the HMB-45. It was discovered by the Dr. Allen Gown and Dr. Arthur Vogel. Since it reacts positively against the melanocytic tumors and not any other tumor, hence demonstrate the specificity and sensitivity. HMB-45 is 92% sensitive for the melanoma and hence is dependable and trustable for the determination of the melanoma tumors. It also reacts positively for the nevus cells besides the fetal melanocytes. It should be kept at 4 degree Celsius and must be used before 2 months period of time in order to keep it intact and maintain its quality.

A small tissue or cell of the targeted area must be collected in order to proceed with the test. While taking the specimen for the test, precaution must be made to avoid spreading of microbial and bacterial infections. The tools to be used must be sterilised properly in order to avoid the spreading of infection Along with tools, personal hygiene and sterilization must be kept in mind. Use of gloves and masks to avoid the risk of the infections is recommended. Lastly, avoid the contact of the reagents with eyes or the mucous membrane Contamination of the reagents must be avoided as well.

Positive staining disease:
Angiomyolipoma
Clear cell sarcoma of the soft tissues
Clear cell sugar tumor
Renal cell carcinoma
Melanocytosis
Pigmented neural tumor
Tuberous sclerosis complex component
Epithelioid sarcoma
Lymphangioleiomyomatosis
PEComa
Negative staining:
Adult melanocyte
Atypical fibroxanthoma
Liposarcoma
Paget disease of breast, vulva and scrotum
Pigmented dermatofibrosarcoma protuberans
Spindle cell melanoma
Desmoplastic or oral mucosal melanoma

Heat Induced Epitope Retrieval (HIER) techniques is the preferred method deparaffinization, epitope retrieval and rehydration. Upon completion, rinse with deionized or distilled water. In case you are using the HRP detection system, place the slides in the peroxide block for at least 10minutes. Omit this step if you are using the AP detection system Now, apply the antibody and incubate for 10 to 30 minutes Rinse it and apply the detection reagent and rinse again Apply enough chromogen and incubate for 10 minutes and rinse again Finally dehydrate and place the coverslip.