Anti-TRP1 antibody [EPR21956]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRP1 antibody [EPR21956] (AB235446)

Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling TRP1 with ab235446 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Cytoplasmic staining in tumor cells of human melanoma (PMID : 22045183; 26068396) is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-TRP1 antibody [EPR21956] (AB235446)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MeWo (human malignant melanoma cell line) cells labeling TRP1 with ab235446 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining in the MeWo cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-TRP1 antibody [EPR21956] (AB235446)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized MeWo (human malignant melanoma cell line) cells labeling TRP1 with ab235446 at 1/500 (right) compared with a Isotype control details (ab172730) (left). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

TRP1 is predominantly expressed in mature melanosomes (PMID : 27073483, 11266471).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRP1 antibody [EPR21956] (AB235446)

Immunohistochemical analysis of paraffin-embedded human skin tissue labeling TRP1 with ab235446 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) ready to use. Positive staining of melanocytes in human skin (PMID : 21562572) is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ready to use.

Heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

• WB

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Western blot - Anti-TRP1 antibody [EPR21956] (AB235446)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times.

Lane 1 : 8 seconds, Lanes 2 & 3 : 3 minutes.

The observed mass is greater than the predicted due to glycosylation.
The molecular mass observed is consistent with what has been described in the literature (PMID : 21324755; PMID : 18042623; 18650849).
The blot with lanes 2 and 3 was developed with a high sensitivity ECL substrate.

All lanes:

Western blot - Anti-TRP1 antibody [EPR21956] (ab235446) at 1/1000 dilution

Lane 1:

MeWo (human malignant melanoma cell line) whole cell lysate at 20 µg

Lane 2:

SK-MEL-2 (human skin malignant melanoma cell line) whole cell lysate at 20 µg

Lane 3:

SK-MEL-28 (human malignant melanoma cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 61 kDa

Observed band size: 75 kDa

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