HMB45 - Pigmented melanoma metastases from LN MART-1 - Recombinant human MART-1 protein
HMB-45 and MART-1 are coexpressed in the majority of melanomas, as well as uniquely expressed in certain cases. Thus, the HMB-45 and MART-1 cocktail is potentially more sensitive than HMB-45 or MART-1 alone. MART-1 is a cocktail of clones DT101 and BC199. The combination of HMB-45 and the MART-1 cocktail make this triple antibody cocktail a first-order pan melanoma screener.
Our Abpromise guarantee covers the use of ab732 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use at an assay dependent concentration. PubMed: 22147606|
|IHC-P||Use at an assay dependent concentration. The staining intensity of this melanoma cocktail is enhanced by performing heat mediated antigen retrieval using a high pH buffer. The HMB-45 portion of this antibody will stain with citrate buffer, pH 6.0; however, data shows the MART-1 portion of this cocktail may be negative unless a high pH buffer is used. We therefore, strongly recommend that you do not use citrate buffer for this antibody cocktail.|
|IHC-Fr||Use at an assay dependent concentration.
For an ABC system, dilute 1/50-1/100 and incubate for 30-60 minutes at RT. It is sometimes difficult to interpret DAB stained melanomas due to endogenous pigment, therefore we recommend you substitute an AEC or a Fast Red substrate protocol.
ab732 staining murine melanoma metastasis tissue sections by IHC-P. Tissue sections were paraformaldehyde fixed and blocked with 0.5% Perkin-Elmer TNB Blocking Buffer for 30 minutes at 25°C. The primary antibody was diluted 1/50 and incubated for 18 hours at 4°C. An Alexa Fluor 488® conjugated goat anti-mouse was used as the secondary
Confocal image shows melanoma in magenta, vessels in red (Collagen Type IV, ab19808), and melanin in cyan. Tissue is perfusion-fixed 15µm cryostat sections mounted on slides (i.e., lightly fixed, but not paraffin embedded).