MA5-13232

antibody from Invitrogen Antibodies

Targeting: PMEL D12S53E, gp100, Pmel17, SI, SIL, SILV

Immunohistochemistry

Antibody data

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Validation data

Product number: MA5-13232 - Provider product page
Provider: Invitrogen Antibodies
Product name: PMEL Monoclonal Antibody (HMB45)
Antibody type: Monoclonal
Antigen: Other
Description: MA5-13232 targets Melanoma (gp100) in IHC (P) and WB applications and shows reactivity with Human samples. The MA5-13232 immunogen is extract of pigmented melanoma metastases from lymph nodes.
Reactivity: Human
Host: Mouse
Isotype: IgG
Antibody clone number: HMB45
Vial size: 500 µL
Concentration: 0.2 mg/mL
Storage: 4° C

PMEL protein structure - MA5-13232 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Identity of the recombinant glutathione-S-transferase (GST)-fusion proteins produced in bacteria. Each protein of interest produced in fusion with GST was detected with a relevant specific antibody. The western blot profiles show a variety of recombinant protein fragments as previously detected by the monoclonal anti-GST antibody, as shown in Figure 2 . (-): bacterial extracts from induced culture of bacteria containing the empty vector pKE-1. (+): bacterial extracts from induced cultures of recombinant bacteria containing the sequence encoding GST-fusion proteins.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 3 Oxysterol quantification in primitive and metastatic melanoma cells and PMEL expression in IGR 37 and IGR 39 melanoma cell line A Absolute quantitation of 24-, 25- and 27-hydroxycholesterol in primitive and metastatic melanoma cells as indicated. ( T -test analysis, *** P < 0.001, N = 3 biological replicates, data are mean +- SD). B Histogram representing R square of correlation analysis between absolute quantitation of 24-, 25- and 27-hydroxycholesterol and label free quantitation of APOE in a cohort of primitive (A375, IPC298) and metastatic (SKMEL5, SKMEL28) melanoma cell lines. C Confocal fluorescence images of anti-HMB45 PMEL antibody signal (green) and DAPI staining (blue) in IGR37 and IGR39. Scale bar is 10 mum. D Western blot on IGR37 and 39 cellular lysates probed with anti-HMB45 PMEL antibody and anti HDAC2 to check the loading of similar amount of total lysates. Source data are available online for this figure.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2 Confocal micrographs of ARPE-19 cells cultured for 4 months. The ARPE-19 cells were plated on laminin-coated Transwell (r) filter membrane at a density of 3 x 10 5 cells/cm 2 and grown for 4 months as outlined in the Methods section. Rabbit anti- zonula occludens-1 (ZO-1), mouse anti-premelanosome protein (PMEL), mouse anti-claudin-2 primary antibodies, and Alexa Fluor secondary antibodies were used to immunostain the ARPE-19 cells at 4 months post-confluency; rhodamine-phalloidin was used to stain actin. A : PMEL immunostaining correlates with the presence of melanosome pigmentation. B : ZO-1 immunostaining shows good junctional development and localization at the cell borders containing a mixture of elongated and polygonal cells. C : Actin immunostaining reveals circumferential actin distribution throughout the height of the cells. D : Claudin-2 immunostaining shows tight junction localization uniformly across the monolayer. E : ZO-1 immunostaining shows good junctional development and localization at the cell borders regardless of the secondary antibody used. F : Claudin-2 immunofluorescent labeling shows colocalization with ZO-1 in the tight junction complexes (merged image of D and E ). Images were taken using an Andor Revolution XD spinning disk confocal microscope using the 40X Plan Fluor oil immersion objective. Scale bar = 15 mum.