Anti-Tyrosinase antibody [EPR10141]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tyrosinase antibody [EPR10141] (AB170905)

Immunohistochemical analysis of paraffin-embedded human skin tissue labelling Tyrosinase with ab170905 at 1/500 (0.662 μg/ml) dilution followed by a ready to use LeicaDS9800 (Bond^™ Polymer Refine Detection). Positive staining on human skin. The section was incubated with ab170905 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Tyrosinase antibody [EPR10141] (AB170905)

ab170905 staining Tyrosinase in A375 (human malignant melanoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 100% methanol and incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used as a nuclear counterstain.

Control : PBS only.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Tyrosinase antibody [EPR10141] (AB170905)

Intracellular Flow Cytometry analysis of A375 (human malignant melanoma) cells labeling with purified ab170905 at 1/30 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Tyrosinase antibody [EPR10141] (AB170905)

Immunohistochemical analysis of paraffin-embedded Human melanoma tissue, labeling Tyrosinase using ab170905 at a 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-Tyrosinase antibody [EPR10141] (AB170905)

All lanes:

Western blot - Anti-Tyrosinase antibody [EPR10141] (ab170905) at 1/1000 dilution

All lanes:

Human melanoma lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 60 kDa

true

• WB

Unknown

Western blot - Anti-Tyrosinase antibody [EPR10141] (AB170905)

Blocking buffer : 5% NFDM/TBST.

Exposure :

Lane 1 : 20 seconds

Lane 2 : 60 seconds

All lanes:

Western blot - Anti-Tyrosinase antibody [EPR10141] (ab170905) at 1/1000 dilution

Lane 1:

SK-MEL-28 (Human malignant melanoma) whole cell lysate at 15 µg

Lane 2:

MeWo (Human malignant melanoma fibroblast) whole cell lysate at 15 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 60 kDa

Observed band size: 70 kDa

false

• WB

Supplier Data

Western blot - Anti-Tyrosinase antibody [EPR10141] (AB170905)

Blocking buffer and concentration : 5% NFDM/TBST Diluting buffer and concentration : 5% NFDM/TBST The expression profile observed is consistent with what has been described in the literature (PMID : 20053998; PMID : 24933607). Negative control : NIH/3T3, RAW264.7 whole cell lysates.

All lanes:

Western blot - Anti-Tyrosinase antibody [EPR10141] (ab170905) at 1/1000 dilution

Lane 1:

B16-F0 (mouse melanoma epithelial-like cell), whole cell lysate at 20 µg

Lane 2:

B16-F10 (mouse melanoma epithelial-like cell), whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Lane 4:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), 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: 60 kDa

Observed band size: 50-80 kDa

false

Exposure time: 59s

• WB

CiteAb

Western blot - Anti-Tyrosinase antibody [EPR10141] (AB170905)

Tyrosinase western blot using anti-Tyrosinase antibody [EPR10141] ab170905. Publication image and figure legend from Jessen, C., Kreß, J. K. C., et al., 2020, Oncogene, PubMed 32978520.

ab170905 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab170905 please see the product overview.

Suppression of differentiation features by NRF2.a Immunoblot of NRF2 and COX2 in UACC-62 NFE2L2 wt and NFE2L2 knockout cells after TNFα treatment (50 ng/ml, 3 d). Vinculin served as loading control. b Corresponding real-time PCR of PTGS2 gene expression, derived from two independent experiments. c Linear regression analysis of MITF and PTGS2 mRNA (n = 472) (Adj R2 = 0.076577 Intercept = 4.4372 Slope = −0.20477 p = 5.7562e-10). The results shown here are based upon data derived from the TCGA dataset Skin Cutaneous Melanoma, and FPKM values were downloaded from www.cbioportal.org. d Protein blot of NRF2 and MITF in UACC-62 cells after knockdown of NFE2L2 for 3 d with two independent siRNAs. MITF is represented by both visible bands (arrows). e Expression changes of pigmentation genes after NFE2L2 knockdown in UACC-62 cells, as detected by RNA sequencing, using the siRNA N2. f Protein blot of TYR and MLANA in UACC-62 cells after knockdown of NFE2L2 for 3 d with two independent siRNAs. g Luciferase assay of UACC-62 cells after MITF induction (100 or 250 ng/ml, 3 d) and co-transfection with 400 ng or 800 ng of pcDNA3.1-NRF2 and a tyrosinase promoter construct for 2 d. Luciferase activity was measured twice in duplicates. h Immunoblot of NRF2 and MITF after doxycycline-dependent NRF2 induction in UACC-62 cells (1000 ng/ml Dox, 3 d). Actin served as loading control. i Immunoblot of MITF and COX2 in UACC-62 cells expressing the Dox-inducible MITF expression vector pSB-MITF. Where indicated, cells were treated with doxycycline (250 ng/ml, 3 d) and H2O2 was added for the last 5 h before harvesting (400 µM). j Corresponding real-time PCR of PTGS2. Data are derived from two independent experiments performed in duplicates. Error bars represent SD.

false