Anti-CD47 antibody [EPR21794]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR21794] (AB218810)

Immunohistochemical analysis of formalin fixed paraffin embedded human non small cell lung carcinoma labelling CD47 with ab218810 at a concentration of 1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab218810 Anti-CD47 antibody [EPR21794] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR21794] (AB218810)

Immunohistochemical analysis of formalin fixed paraffin embedded human ovarian carcinoma labelling CD47 with ab218810 at a concentration of 1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab218810 Anti-CD47 antibody [EPR21794] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR21794] (AB218810)

Immunohistochemical analysis of formalin fixed paraffin embedded human placenta labelling CD47 with ab218810 at a concentration of 1µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab218810 Anti-CD47 antibody [EPR21794] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR21794] (AB218810)

Immunohistochemical analysis of paraffin-embedded human ovary carcinoma tissue labeling CD47 with ab218810 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weakly cytoplasmic staining in human ovary carcinoma (PMID : 7998989). Counterstained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR21794] (AB218810)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling CD47 with ab218810 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous and weak cytoplasmic staining on sinus of human liver sinusoidal endothelial cells (PMID : 7998989). Counterstained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR21794] (AB218810)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling CD47 with ab218810 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Membranous staining on human placenta (PMID : 7998989). Counterstained with hematoxylin. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

• WB

Lab

Western blot - Anti-CD47 antibody [EPR21794] (AB218810)

Lanes 1-4 : Merged signal (red and green). Green - ab218810 observed at 47-52 kDa. Red - loading control ab8245 observed at 36 kDa.

ab218810 Anti-CD47 antibody [EPR21794] was shown to specifically react with CD47 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266324 (knockout cell lysate ab257220) was used. Wild-type and CD47 knockout samples were subjected to SDS-PAGE. ab218810 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CD47 antibody [EPR21794] (ab218810) at 1/500 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

CD47 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human CD47 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-cd47-knockout-hek-293t-cell-line-ab266324'>ab266324</a>)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 35 kDa

Observed band size: 47-52 kDa

false

• WB

Supplier Data

Western blot - Anti-CD47 antibody [EPR21794] (AB218810)

Exposure time : Lane 1 : 26 seconds; Lane 2 : 70 seconds; Lanes 3-4 : 3 minutes; Lane 5 : 3 seconds.

Blocking/Dilution buffer : 5% NFDM/TBST.

We recommend that samples are not boiled after adding loading buffer as this may cause protein aggregates. The molecular mass observed is consistent with the literature (PMID 12393467, PMID 11034562).

Lanes 1 - 4:

Western blot - Anti-CD47 antibody [EPR21794] (ab218810) at 1/5000 dilution

Lane 5:

Western blot - Anti-CD47 antibody [EPR21794] (ab218810) at 1/1000 dilution

Lane 1:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 2:

U937 (human histiocytic lymphoma cell line) whole cell lysate at 20 µg

Lane 3:

A549 (human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

U-2 OS (human bone osteosarcoma epithelial cell line) whole cell lysate at 20 µg

Lane 5:

Human brain 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: 35 kDa

Observed band size: 47-52 kDa

false