Anti-GPNMB antibody [EPR22011-11] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [EPR22011-11] - BSA and Azide free (AB236209)

Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling GPNMB with ab222109 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining in tumor cells of human melanoma. 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab222109).

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GPNMB antibody [EPR22011-11] - BSA and Azide free (AB236209)

Immunohistochemical analysis of paraffin-embedded human colonic carcinoma tissue labeling GPNMB with ab222109 at 1/1000 dilution, followed by aGoat Anti-Rabbit IgG H&L (HRP) Ready to use. Positive staining on stroma cells of human colonic carcinoma. 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab222109).

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-GPNMB antibody [EPR22011-11] - BSA and Azide free (AB236209)

This data was developed using ab222109, the same antibody clone in a different buffer formulation.

ab222109 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 ab222109 overnight at 4 °C at a 1/1000 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 [EPR22011-11] (<a href='/en-us/products/primary-antibodies/gpnmb-antibody-epr22011-11-ab222109'>ab222109</a>) at 1/1000 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 [EPR22011-11] - BSA and Azide free (AB236209)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab222109).

Blocking/Dilution buffer : 5% NFDM/TBST.

The molecular mass observed is consistent with what has been described in the literature (PMID : 19320736).

Negative control : HeLa (PMID : 20056711)

All lanes:

Western blot - Anti-GPNMB antibody [EPR22011-11] - BSA and Azide free (ab236209) at 1/1000 dilution

Lane 1:

SK-MEL-28 (human malignant melanoma) whole cell lysate at 10 µg

Lane 2:

SK-MEL-2 (Human skin malignant melanoma) whole cell lysate at 10 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 10 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: 64 kDa

Observed band size: 120 kDa,95 kDa

false