Anti-GPNMB antibody [SP299]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-GPNMB antibody [SP299] (AB227695)

ab227695 was shown to react with GPNMB in wild-type U-87 MG cells in immunocytochemistry with loss of signal observed in a GPNMB siRNA knockdown cell line. Wild-type and siRNA knockdown cells were mixed and pelleted at a 1 : 1 ratio on coverslips. The cells were fixed with 4% paraformaldehyde (15 min) then permeabilized with 0.1% Triton X-100 (10min) and then blocked with 1x PBS, 0.01% Triton X-100, 5% BSA, 5% NGS. The cells were then incubated with ab227695 at 1/500 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat anti-rabbit secondary antibody to (Alexa Fluor® 555) at 0.5 ?g/ml. Acquisition of the green (wild-type), red (antibody staining) and far-red (siRNA knockdown) channels was performed. Representative grayscale images of the red channel are shown. Wild-type and siRNA knockdown are outlined with yellow and magenta dashed line, respectively. Schematic representation of the mosaic strategy used is shown on the bottom-right panel. Image was acquired with a Zeiss(LSM-880).

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [SP299] (AB227695)

Formalin-fixed paraffin-embedded human reactive lymph node tissue stained for GPNMB using ab227695 at 1/100 dilution in immunohistochemical analysis.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [SP299] (AB227695)

Formalin-fixed paraffin-embedded human tonsil tissue stained for GPNMB using ab227695 at 1/100 dilution in immunohistochemical analysis.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [SP299] (AB227695)

Formalin-fixed paraffin-embedded human lung squamous cell carcinoma tissue stained for GPNMB using ab227695 at 1/100 dilution in immunohistochemical analysis.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-GPNMB antibody [SP299] (AB227695)

Flow Cytometry analysis of A-375 (human malignant melanoma cell line) cells labeling GPNMB with ab227695 at 1/100 dilution (green) compared to a Rabbit IgG negative control (blue).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [SP299] (AB227695)

Formalin-fixed paraffin-embedded human cervical squamous cell carcinoma tissue stained for GPNMB using ab227695 at 1/100 dilution in immunohistochemical analysis.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [SP299] (AB227695)

Formalin-fixed paraffin-embedded human lung tissue stained for GPNMB using ab227695 at 1/100 dilution in immunohistochemical analysis.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [SP299] (AB227695)

Formalin-fixed paraffin-embedded human colon tissue stained for GPNMB using ab227695 at 1/100 dilution in immunohistochemical analysis.

• WB

Lab

Western blot - Anti-GPNMB antibody [SP299] (AB227695)

ab227695 was shown to react with GPNMB in wild-type U-87 MG cells in Western blot with loss of signal observed in a GPNMB siRNA knockdown cell line. Cell lysates from wild-type U-87 MG transfected with either scrambled siRNA or GPNMB siRNA were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab227695 overnight at 4 °C at a 1/500 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-GPNMB antibody [SP299] (ab227695) at 1/500 dilution

Lane 1:

Wild-type U-87 MG transfected with scrambled siRNA control lysate at 60 µg

Lane 2:

U-87 MG transfected with siRNA specifically targeting GPNMB cell lysate at 60 µg

false

• WB

Supplier Data

Western blot - Anti-GPNMB antibody [SP299] (AB227695)

All lanes:

Western blot - Anti-GPNMB antibody [SP299] (ab227695) at 1/100 dilution

All lanes:

A-375 (human malignant melanoma cell line) whole cell lysate

Predicted band size: 64 kDa

false