Anti-CD47 antibody [EPR28770-376]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR28770-376] (AB319049)

Immunohistochemical analysis of paraffin-embedded Mouse lung carcinoma tissue labeling CD47 with ab319049 at 1/5000 (0.101 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse lung carcinoma.
The section was incubated with ab319049 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

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

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 10 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR28770-376] (AB319049)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling CD47 with ab319049 at 1/5000 (0.101 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : very weak staining on mouse skeletal muscle.
The section was incubated with ab319049 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

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

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 10 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD47 antibody [EPR28770-376] (AB319049)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD47 with ab319049 at 1/5000 (0.101 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen.
The section was incubated with ab319049 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument
Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background

Counterstained with Hematoxylin.

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

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 10 mins

• WB

Supplier Data

Western blot - Anti-CD47 antibody [EPR28770-376] (AB319049)

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

CD47 is a glycoprotein of approximately 40-52 kDa and detected as a 25 kDa band after treated with Deglycosylation Peptide : N-glycosidase F (PNGase F).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CD47 antibody [EPR28770-376] (ab319049) at 1/1000 dilution

Lane 1:

Untreated mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse spleen tissue lysate treated with Protein Deglycosylation Peptide:N-glycosidase F (PNGase F) at 20 µg

Lane 3:

Untreated NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 whole cell lysate treated with Protein Deglycosylation Peptide:N-glycosidase F (PNGase F) 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

Observed band size: 40-52 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-CD47 antibody [EPR28770-376] (AB319049)

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

Low expression : skeletal muscle.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 27091975, PMID : 32576895).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CD47 antibody [EPR28770-376] (ab319049) at 1/1000 dilution

Lane 1:

CTLL-2 (mouse T lymphocyte) whole cell lysate at 20 µg

Lane 2:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 3:

B16-F0 (mouse melanoma epithelial-like cell) whole cell lysate at 20 µg

Lane 4:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

Lane 5:

Mouse spleen tissue lysate at 20 µg

Lane 6:

Mouse thymus tissue lysate

Lane 7:

Mouse skeletal muscle tissue 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

Observed band size: 40-52 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-CD47 antibody [EPR28770-376] (AB319049)

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

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-CD47 antibody [EPR28770-376] (ab319049) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

NIH/3T3 transfected with siRNA specifically targeting CD47 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

Observed band size: 285 kDa,36 kDa

false

Exposure time: 37s