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Analysis of MART-1/Melan-A Expression Left Panel - Flow cytometric analysis of MART-1/Melan-A expression by human SK-MEL-5 cells. Cells from the human SK-MEL-5 (Melanoma, ATCC HTB-70) cell line were fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655). The cells were washed and then permeabilized and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with either Purified Mouse IgG1, κ Isotype Control (Cat No. 554121, dashed line histogram) or Purified Mouse Anti-Human MART-1/Melan-A antibody (Cat No. 567886; solid line histogram). The cells were washed and stained with PE Goat Anti-Mouse Ig (Multiple Adsorption) (Cat. No. 550589). The fluorescence histogram showing MART-1/Melan-A expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of intact human SK-MEL-5 cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Right Panel - Immunohistochemical staining of MART-1/Melan-A expressed in human melanoma tissue. Following antigen retrieval with BD Retrievagen A Buffer (Cat. No. 550524), sections from formalin-fixed, paraffin-embedded, human melanoma cell-containing tissue were stained with either Purified Mouse IgG1 κ Isotype Control (Cat. No. 550878, Left Image) or Purified Mouse Anti-Human MART-1/Melan-A antibody (Cat. No. 567886; Right Image), A three-step staining procedure that employs Biotin Goat Anti-Mouse Ig (Cat. No. 550337), Streptavidin HRP (Cat. No. 550946) and DAB (Cat. No. 550880) was used to reveal the primary staining reagents. Counterstaining was with Hematoxylin. Original magnification: 40 X.
BD Pharmingen™ Purified Mouse Anti-Human MART-1/Melan-A
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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- An isotype control should be used at the same concentration as the antibody of interest.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
Companion Products
The A103 monoclonal antibody specifically recognizes Melanoma antigen recognized by T-cells 1 (MART-1) which is also known as Melan-A. This molecule is an ~13 kDa transmembrane protein that is encoded by MLANA and belongs to a melanocyte lineage-specific family of proteins. MART-1 (Melan-A) is found in melanosomes and the endoplasmic reticulum of melanocytes and melanoma cells as well as retinal pigment epithelium. The A103 antibody labels melanomas and other tumors showing melanocytic differentiation. The A103 antibody can also reportedly stain adrenal cortical cells and steroid-producing cells in the ovary and testis. MART-1 (Melan-A) plays a role in melanosome biosynthesis and can serve as a target antigen recognized by cytotoxic T lymphocytes.
Development References (4)
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Chen YT, Stockert E, Jungbluth A, et al. Serological analysis of Melan-A(MART-1), a melanocyte-specific protein homogeneously expressed in human melanomas.. Proc Natl Acad Sci U S A. 1996; 93(12):5915-9. (Immunogen: Immunohistochemistry, Western blot). View Reference
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Coulie PG, Brichard V, Van Pel A, et al. A new gene coding for a differentiation antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas.. J Exp Med. 1994; 180(1):35-42. (Biology). View Reference
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Jungbluth AA, Busam KJ, Gerald WL, et al. A103: An anti-melan-a monoclonal antibody for the detection of malignant melanoma in paraffin-embedded tissues.. Am J Surg Pathol. 1998; 22(5):595-602. (Clone-specific). View Reference
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Kawakami Y, Eliyahu S, Delgado CH, et al. Cloning of the gene coding for a shared human melanoma antigen recognized by autologous T cells infiltrating into tumor.. Proc Natl Acad Sci U S A. 1994; 91(9):3515-9. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.