Anti-EMR1/ADGRE1 (F4/80) antibody [RM1302]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EMR1/ADGRE1 (F4/80) antibody [RM1302] (AB323648)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling EMR1/ADGRE1 (F4/80) with ab323648 at 1/500 (0.946 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on Kupffer cells of human liver.
The section was incubated with ab323648 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 was performed 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-EMR1/ADGRE1 (F4/80) antibody [RM1302] (AB323648)

Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling EMR1/ADGRE1 (F4/80) with ab323648 at 1/500 (0.946 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of human lung cancer.
The section was incubated with ab323648 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 was performed 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-EMR1/ADGRE1 (F4/80) antibody [RM1302] (AB323648)

Immunohistochemical analysis of paraffin-embedded Human skeletal muscle tissue labeling EMR1/ADGRE1 (F4/80) with ab323648 at 1/500 (0.946 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : no staining on human skeletal muscle.
The section was incubated with ab323648 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 was performed 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-EMR1/ADGRE1 (F4/80) antibody [RM1302] (AB323648)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling EMR1/ADGRE1 (F4/80) with ab323648 at 1/2000 (0.237 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : positive staining on immune cells of mouse skeletal muscle.
The section was incubated with ab323648 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 was performed 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-EMR1/ADGRE1 (F4/80) antibody [RM1302] (AB323648)

Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling EMR1/ADGRE1 (F4/80) with ab323648 at 1/2000 (0.237 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on rat spleen.
The section was incubated with ab323648 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 was performed 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-EMR1/ADGRE1 (F4/80) antibody [RM1302] (AB323648)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling EMR1/ADGRE1 (F4/80) with ab323648 at 1/2000 (0.237 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen.
The section was incubated with ab323648 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt

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Flow Cytometry - Anti-EMR1/ADGRE1 (F4/80) antibody [RM1302] (AB323648)

Flow cytometric analysis of mouse PBMC cells labelling EMR1/ADGRE1 (F4/80) with ab323648 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

mouse PBMC are co-stained with CD11b conjugated BV510.
Gated on viable cell.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EMR1/ADGRE1 (F4/80) antibody [RM1302] (AB323648)

Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling EMR1/ADGRE1 (F4/80) with ab323648 at 1/2000 (0.237 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression : positive staining on immune cells of rat skeletal muscle.
The section was incubated with ab323648 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-EMR1/ADGRE1 (F4/80) antibody [RM1302] (AB323648)

Flow cytometric analysis of mouse PBMC cells labelling EMR1/ADGRE1 (F4/80) with ab323648 at 1/50 dilution (1ug) / Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control.

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

mouse PBMC are co-stained with CD3 conjugated Alexa Fluor®647.
Gated on viable cell.

• IP

Supplier Data

Immunoprecipitation - Anti-EMR1/ADGRE1 (F4/80) antibody [RM1302] (AB323648)

EMR1/ADGRE1 (F4/80) was immunoprecipitated from 0.35 mg Mouse lymph node tissue lysate with ab323648 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab323648 at 1/1000 dilution.

Blocking and dilution buffer and concentration : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-EMR1/ADGRE1 (F4/80) antibody [RM1302] (ab323648) at 1/1000 dilution

Lane 1:

Mouse lymph node tissue lysate at 10 µg

Lane 2:

ab323648 at 1/30 IP in Mouse lymph node tissue lysate at 10 µg

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab323648 in Mouse lymph node tissue lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 90-160 kDa

false

Exposure time: 154s

• WB

Supplier Data

Western blot - Anti-EMR1/ADGRE1 (F4/80) antibody [RM1302] (AB323648)

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

Low expression : cerebellum and skeletal muscle.

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

All lanes:

Western blot - Anti-EMR1/ADGRE1 (F4/80) antibody [RM1302] (ab323648) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Lane 4:

Human spleen tissue lysate at 20 µg

Lane 5:

Human lymph node 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 1/20000 dilution

Observed band size: 120 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-EMR1/ADGRE1 (F4/80) antibody [RM1302] (AB323648)

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

Negative control : NIH/3T3 (PMID : 8550607).

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-EMR1/ADGRE1 (F4/80) antibody [RM1302] (ab323648) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage ) 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 1/20000 dilution

Observed band size: 90-160 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-EMR1/ADGRE1 (F4/80) antibody [RM1302] (AB323648)

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

F4/80 (ADGRE1) is a glycoprotein of approximately 120~160 kDa and detected as a 70-90 kDa band after treated with Protein Deglycosylation MIX II. (or after Protein Deglycosylation Mix II treatment) (PMID : 8550607)

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

All lanes:

Western blot - Anti-EMR1/ADGRE1 (F4/80) antibody [RM1302] (ab323648) at 1/1000 dilution

Lane 1:

Untreated RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 15 µg

Lane 2:

RAW 264.7 lysate treated with Protein Deglycosylation Mix II at 15 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 1/20000 dilution

Observed band size: 70-160 kDa,36 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-EMR1/ADGRE1 (F4/80) antibody [RM1302] (AB323648)

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

Low expression : brain and skeletal muscle.

The identity of the lower MW band at approximately 25 kDa (in lane 1-4) is unknown.

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-EMR1/ADGRE1 (F4/80) antibody [RM1302] (ab323648) at 1/1000 dilution

Lane 1:

Mouse spleen tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 5:

Rat spleen tissue lysate at 20 µg

Lane 6:

Rat liver tissue lysate at 20 µg

Lane 7:

Rat brain tissue lysate at 20 µg

Lane 8:

Rat 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 1/20000 dilution

Observed band size: 90-160 kDa,36 kDa

false

Exposure time: 180s