Melanoma gp100 Recombinant Rabbit Monoclonal Antibody [JE42-47]
cat.: HA721719
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse
Applications: WB, IHC-P, IF-Cell
Clonality: Monoclonal
Clone number: JE42-47
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 70 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human Melanoma gp100 aa 617-661 / 661.
Positive control: SK-MEL-28 cell lysates, human melanoma tissue, mouse testis tissue, SK-MEL-28.
Subcellular location: Endoplasmic reticulum membrane, Golgi apparatus, cis-Golgi network membrane, Endosome, multivesicular body, Melanosome, Extracellular vesicle, Secreted.
Recommended Dilutions:
  WB
  IHC-P
  IF-Cell
1:1,000
1:1,000
1:100
Uniprot #: SwissProt: P40967 Human | Q60696 Mouse
Alternative names: 95 kDa melanocyte specific secreted glycoprotein 95 kDa melanocyte-specific secreted glycoprotein D12S53E gp100 M-beta ME20 ME20 M/ME20 S ME20-M ME20-S ME20M ME20M/ME20S ME20S Melanocyte lineage specific antigen GP100 Melanocyte protein mel 17 Melanocyte protein Pmel 17 Melanocyte protein Pmel 17 precursor Melanocytes lineage-specific antigen GP100 Melanoma associated ME20 antigen Melanoma gp100 Melanoma-associated ME20 antigen Melanosomal matrix protein 17 Melanosomal matrix protein17 P1 p100 p26 PMEL 17 PMEL PMEL_HUMAN PMEL17 Premelanosome protein Secreted melanoma-associated ME20 antigen SI SIL SILV Silver (mouse homolog) like Silver homolog Silver locus protein homolog Silver, mouse, homolog of
Images
HA721719_1.jpg Fig1: Western blot analysis of Melanoma gp100 on SK-MEL-28 cell lysates with Rabbit anti-Melanoma gp100 antibody (HA721719) at 1/1,000 dilution.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 70 kDa
Observed band size: 100 kDa

Exposure time: 2 minutes;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721719) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.
HA721719_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human melanoma tissue with Rabbit anti-Melanoma gp100 antibody (HA721719) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721719) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721719_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Melanoma gp100 antibody (HA721719) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA721719) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA721719_4.jpg Fig4: Immunocytochemistry analysis of SK-MEL-28 cells labeling Melanoma gp100 with Rabbit anti-Melanoma gp100 antibody (HA721719) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Melanoma gp100 antibody (HA721719) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.