Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling Integrin beta 3 with ab179473 at 1/800 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Positive staining on platelets (PMID: 25356112) of Human spleen is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

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

All lanes : Anti-Integrin beta 3 antibody [ERP17507] (ab179473) at 1/500 dilution

Lane 1 : U-87MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 2 : HEL (Human Erythroleukemia erythroblast) whole cell lysates with 5% NFDM/TBST
Lane 3 : LnCaP (Human prostate carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 4 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 5 : HUVEC (Human umbilical vein endothelial cell) whole cell lysates with 5% NFDM/TBST
Lane 6 : PC-3 (Human prostate adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 7 : C2C12 (Mouse myoblasts myoblast) whole cell lysates with 5% NFDM/TBST
Lane 8 : Raw264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates with 5% NFDM/TBST
Lane 9 : Mouse brain lysates with 5% NFDM/TBST

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 87 kDa
Observed band size: 110 kDa why is the actual band size different from the predicted?


Exposure time: 10 seconds

LnCaP, HeLa and PC-3 are low expression controls. This product doesn’t react with Ms

Purified ab179473 at 1/30 dilution (2µg) immunoprecipitating Integrin beta 3 in HUVEC whole cell lysate.
Lane 1 (input): HUVEC (human umbilical vein endothelial cell) whole cell lysate 10 µg
Lane 2 (+): ab179473 + HUVEC whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab179473 in HUVEC whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 110 kDa

Anti-Integrin beta 3 antibody [ERP17507] (ab179473) at 1000 cells + HEL 92.1.7 (Human bone marrow erythroleukemia cell line) whole cell lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 87 kDa
Observed band size: 97 kDa why is the actual band size different from the predicted?


Exposure time: 5 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.

The molecular weight observed is consistent with the description in the literature (PMID: 1710517).

Anti-Integrin beta 3 antibody [ERP17507] (ab179473) at 1/5000 dilution + Human plasma at 10 µg

Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 87 kDa
Observed band size: 97 kDa why is the actual band size different from the predicted?


Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The molecular weight observed is consistent with the description in the literature (PMID: 1710517).

Immunohistochemical analysis of paraffin-embedded Human osteosarcomas tissue labeling Integrin beta 3 with ab179473 at 1/800 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane staining on cells of Human osteosarcomas is observed; several multinucleated osteoclasts are positively stained (PMID: 15692727; PMID: 25534583).

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

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

Immunohistochemical analysis of paraffin-embedded Human thyroid cancer tissue labeling Integrin beta 3 with ab179473 at 1/800 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Membrane or cytoplasmic staining on tumor cells of Human thyroid cancer is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

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