MLANA/MART-1 Antibody [D21A21] | Primary Antibodies

Application: Reactivity: Human

Lane 1: SK-MEL-28

Usage Information

Dilution
WB1:1000 IP1:50 IHC1:200 IF1:800 FCM1:50-1:200

Application
WB, IP, IHC, IF, FCM

Reactivity
Human

Source
Rabbit Monoclonal Antibody

Storage Buffer
PBS, pH 7.2+50% Glycerol+0.05% BSA+0.01% NaN3

Storage (from the date of receipt)
-20°C (avoid freeze-thaw cycles), 2 years

Predicted MW
19 kDa

Datasheet & SDS

Biological Description

Specificity
MLANA/MART-1 Antibody [D21A21] detects endogenous levels of total MLANA/MART-1 protein.

Clone
D21A21

Synonym(s)
Melanoma antigen recognized by T-cells 1; MART-1; MLANA

Background
MLANA/MART-1, recognized as a melanocyte-lineage specific transmembrane glycoprotein, functions within the specialized compartment of melanosome biogenesis alongside proteins like PMEL17 and GPR143 to orchestrate pigment organelle maturation. The protein adopts a type I topology with a single transmembrane domain that anchors it across ER, Golgi, and melanosomal membranes, featuring an N-terminal lumenal ectodomain rich in serine/threonine residues for glycosylation and a short cytoplasmic tail that engages sorting signals for trafficking. MLANA physically complexes with PMEL17 in the trans-Golgi network, stabilizing its immature form and promoting proteolytic cleavage into functional Mα and Mβ fragments essential for stage II melanosome fibril assembly, while direct binding to GPR143 stabilizes this GPCR to maintain early melanosome identity and composition during acidification. Conformational epitopes exposed on the ectodomain direct retrograde trafficking to ER-endosomal vesicles via AP-1/3 clathrin adaptors, positioning MLANA for MHC class I loading, where A27L or AAGIGILTV peptides elicit CD8+ T cell cytolysis through TCR-MHC recognition. Transcriptional regulation by MITF binds conserved E-boxes in the MLANA promoter, synchronizing expression with pigmentation genes like TYR and TYRP1 during melanocyte differentiation. Restricted to melanocytes, retinal pigment epithelium, and nevi, persistent surface exposure in melanoma facilitates immune surveillance, with loss correlating to immune escape and dedifferentiation. Aberrant glycosylation alters trafficking kinetics, enhancing metastatic dissemination while retaining immunogenicity for adoptive T cell therapies.

Background

MLANA/MART-1, recognized as a melanocyte-lineage specific transmembrane glycoprotein, functions within the specialized compartment of melanosome biogenesis alongside proteins like PMEL17 and GPR143 to orchestrate pigment organelle maturation. The protein adopts a type I topology with a single transmembrane domain that anchors it across ER, Golgi, and melanosomal membranes, featuring an N-terminal lumenal ectodomain rich in serine/threonine residues for glycosylation and a short cytoplasmic tail that engages sorting signals for trafficking. MLANA physically complexes with PMEL17 in the trans-Golgi network, stabilizing its immature form and promoting proteolytic cleavage into functional Mα and Mβ fragments essential for stage II melanosome fibril assembly, while direct binding to GPR143 stabilizes this GPCR to maintain early melanosome identity and composition during acidification. Conformational epitopes exposed on the ectodomain direct retrograde trafficking to ER-endosomal vesicles via AP-1/3 clathrin adaptors, positioning MLANA for MHC class I loading, where A27L or AAGIGILTV peptides elicit CD8+ T cell cytolysis through TCR-MHC recognition. Transcriptional regulation by MITF binds conserved E-boxes in the MLANA promoter, synchronizing expression with pigmentation genes like TYR and TYRP1 during melanocyte differentiation. Restricted to melanocytes, retinal pigment epithelium, and nevi, persistent surface exposure in melanoma facilitates immune surveillance, with loss correlating to immune escape and dedifferentiation. Aberrant glycosylation alters trafficking kinetics, enhancing metastatic dissemination while retaining immunogenicity for adoptive T cell therapies.

References
https://pubmed.ncbi.nlm.nih.gov/12819038/ https://pubmed.ncbi.nlm.nih.gov/15695812/

Reference Table for Selecting PVDF Membrane Pore Size Specifications

Protein Size (kDa)	PVDF Transfer Membrane Pore Size (μm)
< 10	0.22
10-20	0.22
20-30	0.45
30-45	0.45
45-70	0.45
80-100	0.45
100-140	0.45
> 140	0.45

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Protein Size (kDa)	Separating Gel Percentage
4-40	20%
12-45	15%
10-70	12.5%
15-100	10%
25-100	8%
60-210	5%