Anti-TRP2/DCT antibody [EPR21986]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRP2/DCT antibody [EPR21986] (AB221144)

Immunohistochemical analysis of paraffin-embedded human skin tissue labeling TRP2/DCT with ab221144 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in melanocytes of human skin (PMID : 12535195). 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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-TRP2/DCT antibody [EPR21986] (AB221144)

Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling TRP2/DCT with ab221144 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in human melanoma (PMID : 24475287). 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.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• mIHC

Supplier Data

Multiplex immunohistochemistry - Anti-TRP2/DCT antibody [EPR21986] (AB221144)

Panel A : merged staining of anti-Desmoglein 1/DSG1 (gray; Opal™690), anti-Langerin (green; Opal™520) and anti-TRP2/DCT (red; Opal™570) on human skin. Panel B : anti-TRP2/DCT stained on melanocytes. Panel C : anti-Langerin stained on Langerhans cells. Panel D : anti-Desmoglein 1/DSG1 stained on epidermis. The section was incubated in three rounds of staining : in the order of ab236259, ab283686, and ab221144 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• IP

Supplier Data

Immunoprecipitation - Anti-TRP2/DCT antibody [EPR21986] (AB221144)

TRP2/DCT was immunoprecipitated from 0.35 mg SK-MEL-2 (human skin malignant melanoma cell line) whole cell lysate with ab221144 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221144 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : SK-MEL-2 whole cell lysate 10 μg (Input).

Lane 2 : ab221144 IP in SK-MEL-2 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab221144 in SK-MEL-2 whole cell lysate.

Blocking/Dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 5 seconds.

All lanes:

Immunoprecipitation - Anti-TRP2/DCT antibody [EPR21986] (ab221144)

Predicted band size: 59 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-TRP2/DCT antibody [EPR21986] (AB221144)

TRP2/DCT was immunoprecipitated from 0.35 mg B16-F0 (mouse melanoma epithelial cell-like cell line) whole cell lysate with ab221144 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221144 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : B16-F0 whole cell lysate 10 μg (Input).

Lane 2 : ab221144 IP in B16-F0 whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab221144 in B16-F0 whole cell lysate.

Blocking/Dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 41 seconds.

~85KD band is the glycosylated form (PMID : 12719423)

All lanes:

Immunoprecipitation - Anti-TRP2/DCT antibody [EPR21986] (ab221144)

Predicted band size: 59 kDa

false

• WB

Supplier Data

Western blot - Anti-TRP2/DCT antibody [EPR21986] (AB221144)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure time : Lane 1 : 5 seconds; Lane 2 : 15 seconds; Lane 3 : 26 seconds; Lane 4 : 3 minutes.

The molecular mass observed is consistent with what has been described in the literature (PMID : 12719423, PMID : 25009701)

All lanes:

Western blot - Anti-TRP2/DCT antibody [EPR21986] (ab221144) 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

Lane 4:

F9 (mouse embryonic testicular cancer cell line) whole cell lysate at 20 µg

Secondary

Lane 1:

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

Lanes 2 - 4:

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 1/20000 dilution

Predicted band size: 59 kDa

Observed band size: 69 kDa,85 kDa

false