Anti-CSF-1-R antibody [RM2042]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CSF-1-R antibody [RM2042] (AB315490)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CSF-1-R with ab315490 at 1/2000 (0.248 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on human tonsil. The section was incubated with ab315490 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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-CSF-1-R antibody [RM2042] (AB315490)

Immunohistochemical analysis of paraffin-embedded Human pancreas carcinoma tissue labeling CSF-1-R with ab315490 at 1/2000 (0.248 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human pancreas carcinoma. The section was incubated with ab315490 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-CSF-1-R antibody [RM2042] (AB315490)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling CSF-1-R with ab315490 at 1/2000 (0.248 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on stromal cells of human colon. The section was incubated with ab315490 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CSF-1-R antibody [RM2042] (AB315490)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized THP-1 (human monocytic leukemia monocyte) cells labelling CSF-1-R with ab315490 at 1/100 (4.95 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/mL) dilution (Green).

Confocal image showing mainly membranous staining in THP-1 cell line.
Negative control : Raji.
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) 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 (2 ug/mL) dilution.

• IP

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Immunoprecipitation - Anti-CSF-1-R antibody [RM2042] (AB315490)

CSF-1-R was immunoprecipitated from 0.35 mg THP-1 (human monocytic leukemia monocyte) whole cell lysate with ab315490 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab315490 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

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

Lysates were freshly made and used for Immunoprecipitation immediately to minimize protein degradation.

All lanes:

Immunoprecipitation - Anti-CSF-1-R antibody [RM2042] (ab315490) at 1/1000 dilution

Lane 1:

THP-1 (human monocytic leukemia monocyte) whole cell lysate

Lane 2:

ab315490 at 1/30 IP in THP-1 (human monocytic leukemia monocyte) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab315490 in THP-1 whole cell lysate

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

false

Exposure time: 180s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CSF-1-R antibody [RM2042] (AB315490)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CSF-1-R with ab315490 at 1/100 (4.95 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen. The section was incubated with ab315490 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CSF-1-R antibody [RM2042] (AB315490)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling CSF-1-R with ab315490 at 1/100 (4.95 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/mL) dilution (Green).

Confocal image showing membranous and cytoplasmic staining in RAW 264.7 cell line.
Negative control : C2C12
Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) 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 (2 ug/mL) dilution.

• WB

Supplier Data

Western blot - Anti-CSF-1-R antibody [RM2042] (AB315490)

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

Negative control : Raji, HDLM-2 (PMID : 26066800).

The expression molecular weight observed is consistent with what has been described in the literature (PMID : 14673177).

In lanes 1-3, the lysates were stored at -80°C prior to Western Blotting. The bands beneath the target band (140 kDa) are likey to be degradation products. In lanes 4-5, the lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.

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

All lanes:

Western blot - Anti-CSF-1-R antibody [RM2042] (ab315490) at 1/1000 dilution

Lanes 1 and 4:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lanes 2 and 5:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

HDLM-2 (human Hodgkin lymphoma) 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: 190 kDa,140 kDa,36 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-CSF-1-R antibody [RM2042] (AB315490)

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

Negative control : C2C12.

The expression molecular weight observed is consistent with what has been described in the literature (PMID : 14673177).

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

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

All lanes:

Western blot - Anti-CSF-1-R antibody [RM2042] (ab315490) at 1/1000 dilution

Lane 1:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 2:

C2C12 (mouse myoblasts myoblast) 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: 190 kDa,140 kDa,36 kDa

false

Exposure time: 26s