Anti-CSF-1-R antibody [EPR23529-26]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-CSF-1-R antibody [EPR23529-26] (AB254357)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized RAW264.7 cells labelling CSF-1-R with ab254357 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in RAW 264.7 cell line ab195889 Anti-alpha Tubulin antibody [DM1A] - 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 ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CSF-1-R antibody [EPR23529-26] (AB254357)

Immunohistochemical analysis of paraffin-embedded Mouse liver tissue labeling CSF-1-R with ab254357 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on Kupffer cells in mouse liver. The section was incubated with ab254357 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 with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CSF-1-R antibody [EPR23529-26] (AB254357)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CSF-1-R with ab254357 at 1/100 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on macrophages in mouse spleen. The section was incubated with ab254357 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 with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins

• IP

Unknown

Immunoprecipitation - Anti-CSF-1-R antibody [EPR23529-26] (AB254357)

CSF-1-R was immunoprecipitated from 0.35 mg RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate with ab254357 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254357 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 ug

Lane 2 : ab254357 IP in RAW 264.7 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab254357 in RAW 264.7 whole cell lysate

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

Exposure time : 128 seconds.

All lanes:

Immunoprecipitation - Anti-CSF-1-R antibody [EPR23529-26] (ab254357)

Predicted band size: 108 kDa

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• WB

Supplier Data

Western blot - Anti-CSF-1-R antibody [EPR23529-26] (AB254357)

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times.

Lanes 1-2 : 70 seconds; Lanes 3-4 : 3 minutes.

The expression profile and molecular weight observed is consistent with what has been described in the literature (PMID : 14673177)
This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Western blot - Anti-CSF-1-R antibody [EPR23529-26] (ab254357) 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:

J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 µg

Lane 4:

Mouse placenta 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

Predicted band size: 108 kDa

false