Anti-CD47 antibody [RM1014]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [RM1014] (AB284132)

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma tissue labelling CD47 with ab284132 at 1/500 (0.924 μg/ml) followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection) was used. Positive staining on human ovarian carcinoma. The section was incubated with ab284132 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD47 antibody [RM1014] (AB284132)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized Jurkat cells labelling CD47 with ab284132 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in Jurkat cell line. Negative control : HepG2 (PMID : 25721088). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [RM1014] (AB284132)

Immunohistochemical analysis of paraffin-embedded human spleen tissue labelling CD47 with ab284132 at 1/500 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human spleen. The section was incubated with ab284132 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt

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Flow Cytometry - Anti-CD47 antibody [RM1014] (AB284132)

Flow cytometric analysis of HepG2 (Human hepatocellular carcinoma epithelial cell, Left) / U-937 (Human histiocytic lymphoma monocyte, Right) cells labelling CD47 with ab284132 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Low expression control : HepG2 (PMID : 25721088, 30415063). Gated on viable cells.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [RM1014] (AB284132)

Immunohistochemical analysis of paraffin-embedded human ovarian carcin tissue labelling CD47 with ab284132 at 1/500 followed by a ready to use LeicaDS9800 (BOND™Polymer Refine Detection) was used. Positive staining on human ovarian carcinoma. The section was incubated with ab284132 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [RM1014] (AB284132)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labelling CD47 with ab284132 at 1/500 followed by a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection). Positive staining on human placenta. The section was incubated with ab284132 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD47 antibody [RM1014] (AB284132)

Flow cytometric analysis of HepG2 (Human hepatocellular carcinoma epithelial cell, Left) / Jurkat (Human T cell leukemia T lymphocyte, Right) cells labelling CD47 with ab284132 at 1/500 dilution (0.1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Low expression control : HepG2 (PMID : 25721088, 30415063). Gated on viable cells.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [RM1014] (AB284132)

Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labelling CD47 with ab284132 at 1/500 followed by a ready to use LeicaDS9800 (BOND™Polymer Refine Detection). Positive staining on human breast cancer (PMID : 31528120). The section was incubated with ab284132 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• WB

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Western blot - Anti-CD47 antibody [RM1014] (AB284132)

Blocking and diluting buffer and concentration was 5% NFDM/TBST.

We recommend that samples are not boiled after adding loading buffer as this may cause protein aggregates.

All lanes:

Western blot - Anti-CD47 antibody [RM1014] (ab284132) at 1/1000 dilution

Lane 1:

Human liver lysate at 20 µg

Lane 2:

Human ovary cancer 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

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-CD47 antibody [RM1014] (AB284132)

Blocking and diluting buffer and concentration was 5% NFDM/TBST.

Low expression control : HepG2 (PMID : 25721088, 30415063).

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).

All lanes:

Western blot - Anti-CD47 antibody [RM1014] (ab284132) at 1/1000 dilution

Lane 1:

Jurkat (Human T cell leukemia T lymphocyte) whole cell lysate 20ug

Lane 2:

U-937 (Human histiocytic lymphoma monocyte) whole cell lysate 20ug

Lane 3:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 20ug

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

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-CD47 antibody [RM1014] (AB284132)

Blocking and diluting buffer and concentration was 5% NFDM/TBST.

We recommend that samples are not boiled after adding loading buffer as this may cause protein aggregates.

All lanes:

Western blot - Anti-CD47 antibody [RM1014] (ab284132) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (Human embryonic kidney epithelial cell, ab255449) whole cell lysate at 20 µg

Lane 2:

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

false

Exposure time: 37s