Anti-SILV/PMEL Antibody Picoband™

Product Name

Anti-SILV/PMEL Antibody Picoband™

View all PMEL17/SILV Antibodies

SKU/Catalog Number

A01262-3

Size

100 µg/vial

Form

Lyophilized

Description

Boster Bio Anti-SILV/PMEL Antibody Picoband™ catalog # A01262-3. Tested in WB, IHC, FCM, ELISA applications. This antibody reacts with Human, Mouse, Rat.

Storage & Handling

At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.

Cite This Product

Anti-SILV/PMEL Antibody Picoband™ (Boster Biological Technology, Pleasanton CA, USA, Catalog # A01262-3)

Host

Rabbit

Contents

Each vial contains 4 mg Trehalose, 0.9 mg NaCl, 0.2 mg Na2HPO4.

Clonality

Polyclonal

Clone Number

1B9

Isotype

IgG

Immunogen

E.coli-derived human SILV/PMEL recombinant protein (Position: H182-Q5565). Human PMEL shares 73.9% amino acid (aa) sequence identity with mouse PMEL.

*Blocking peptide can be purchased. Costs vary based on immunogen length. Contact us for pricing.

Cross-reactivity

No cross reactivity with other proteins.

Reactive Species

A01262-3 is reactive to PMEL in Human, Mouse, Rat

Applications

A01262-3 is guaranteed for ELISA, Flow Cytometry, IHC, WB Boster Guarantee

Observed Molecular Weight

80 kDa

Calculated molecular weight

70.255kDa

Background of PMEL17/SILV

This gene is mapped to 12q13.2. It encodes a melanocyte-specific type I transmembrane glycoprotein. The encoded protein is enriched in melanosomes, which are the melanin-producing organelles in melanocytes, and plays an essential role in the structural organization of premelanosomes. This protein is involved in generating internal matrix fibers that define the transition from Stage I to Stage II melanosomes. This protein undergoes a complex pattern of prosttranslational processing and modification that is essential to the proper functioning of the protein. A secreted form of this protein that is released by proteolytic ectodomain shedding may be used as a melanoma-specific serum marker. Alternate splicing results in multiple transcript variants.

Antibody Validation

Boster validates all antibodies on WB, IHC, ICC, Immunofluorescence, and ELISA with known positive control and negative samples to ensure specificity and high affinity, including thorough antibody incubations.

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Reconsitution

Adding 0.2 ml of distilled water will yield a concentration of 500 µg/ml.

Assay Dilutions Recommendation

The recommendations below provide a starting point for assay optimization. The actual working concentration varies and should be decided by the user.

Western blot, 0.25-0.5 µg/ml, Human, Mouse, Rat
Immunohistochemistry, 2-5 µg/ml, Human
Flow Cytometry (Fixed), 1-3 µg/1x10⁶ cells, Human
ELISA, 0.1-0.5 µg/ml, Human

Validation Images & Assay Conditions

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Figure 1. Western blot analysis of SILV/PMEL using anti-SILV/PMEL antibody (A01262-3).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human A2058 whole cell lysates,
Lane 2: rat testis tissue lysates,
Lane 3: rat C6 whole cell lysates,
Lane 4: mouse testis tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-SILV/PMEL antigen affinity purified polyclonal antibody (Catalog # A01262-3) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for SILV/PMEL at approximately 80 kDa. The expected band size for SILV/PMEL is at 70 kDa.

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Figure 2. IHC analysis of SILV/PMEL using anti-SILV/PMEL antibody (A01262-3).
SILV/PMEL was detected in a paraffin-embedded section of human melanoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-SILV/PMEL Antibody (A01262-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 3. IHC analysis of SILV/PMEL using anti-SILV/PMEL antibody (A01262-3).
SILV/PMEL was detected in a paraffin-embedded section of human melanoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-SILV/PMEL Antibody (A01262-3) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

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Figure 4. Flow Cytometry analysis of U20S cells using anti-SILV/PMEL antibody (A01262-3).
Overlay histogram showing U20S cells stained with A01262-3 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-SILV/PMEL Antibody (A01262-3, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

Gene/Protein Information For PMEL (Source: Uniprot.org, NCBI)

Alternative Names

D12S53EP1; gp100; ME20; ME20-M; melanocyte protein mel 17; Melanocyte protein Pmel 17; Melanocytes lineage-specific antigen GP100; Melanoma-associated ME20 antigen; melanosomal matrix protein17; PMEL17P100; premelanosome proteinME20M; SI; SIL; silver (mouse homolog) like; silver homolog (mouse); Silver locus protein homolog; silver, mouse, homolog of; SILVPmel17 PMEL D12S53E, ME20, ME20-M, ME20M, P1, P10017, SI, SIL, SILV, gp100, PMEL premelanosome protein melanocyte protein PMEL|melanocyte protein Pmel 17|melanocyte protein mel 17|melanocytes lineage-specific antigen GP100|melanoma-associated ME20 antigen|melanosomal matrix protein17|silver locus protein homolog|silver, mouse, homolog of

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In this infographic, you will see the following information for PMEL: database IDs, superfamily, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post-translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact [email protected].

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