Anti-CX3CR1 antibody [EPR24267-2]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CX3CR1 antibody [EPR24267-2] (AB308613)

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscl tissue labelling CX3CR1 with ab308613 at 1/1000 (0.479 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on mouse skeletal muscle. The section was incubated with ab308613 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CX3CR1 antibody [EPR24267-2] (AB308613)

Immunohistochemical analysis of paraffin-embedded Mouse lung tissue labelling CX3CR1 with ab308613 at 1/1000 (0.479 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on immune cells of mouse lung. The section was incubated with ab308613 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CX3CR1 antibody [EPR24267-2] (AB308613)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling CX3CR1 with ab308613 at 1/1000 (0.479 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on microglial cells of mouse cerebrum. The section was incubated with ab308613 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CX3CR1 antibody [EPR24267-2] (AB308613)

Immunohistochemical analysis of paraffin-embedded Mouse breast cancer tissue labelling CX3CR1 with ab308613 at 1/1000 (0.479 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on immune cells of mouse breast cancer. The section was incubated with ab308613 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrumentIncubate 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 Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CX3CR1 antibody [EPR24267-2] (AB308613)

Immunohistochemical analysis of paraffin-embedded (A) Cerebrum tissue from wild-type C57BL/6J mice and (B) Cerebrum tissue from Cx3cr1 knockout mice tissue labeling CX3CR1 with ab308613 at 1/2000 dilution.

Positive staining on (A) Cerebrum tissue from wild-type C57BL/6J mice, and no staining on (B) Cerebrum tissue from Cx3cr1 knockout mice.

The primary antibody was incubated for 30 mins at room temperature. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins.

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Cx3cr1-KO homozygous mice (Strain ID : T006121).

• IP

Supplier Data

Immunoprecipitation - Anti-CX3CR1 antibody [EPR24267-2] (AB308613)

CX3CR1 was immunoprecipitated from 0.35 mg Mouse spinal cord tissue lysate with ab308613 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab308613 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : Mouse spinal cord tissue lysate Lane 2 : abAB308613 IP in Mouse spinal cord tissue lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab308613 in Mouse spinal cord tissue lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 41 seconds Input is non-boiled as boiling may cause protein aggregates.

All lanes:

Immunoprecipitation - Anti-CX3CR1 antibody [EPR24267-2] (ab308613) at 1/30 dilution

All lanes:

Mouse spinal cord 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: 36 kDa

false

Exposure time: 41s

• WB

Supplier Data

Western blot - Anti-CX3CR1 antibody [EPR24267-2] (AB308613)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : Skeletal muscle (PMID : 12881312). Lanes 4-5 of the blot were freshly made and used for Western blotting immediately to minimize protein degradation. Samples are non-boiled as boiling may cause protein aggregation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-CX3CR1 antibody [EPR24267-2] (ab308613) at 1/1000 dilution

Lanes 1 and 4:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse spinal cord tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 5:

Rat brain 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: 40 kDa

Observed band size: 36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-CX3CR1 antibody [EPR24267-2] (AB308613)

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

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

Samples are non-boiled as boiling may cause protein aggregation. The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and Cx3cr1-KO homozygous mice (Strain ID : T006121).

All lanes:

Western blot - Anti-CX3CR1 antibody [EPR24267-2] (ab308613) at 1/1000 dilution

Lane 1:

Wild-type mouse brain tissue lysate (male) at 20 µg

Lane 2:

Cx3cr1 knockout mouse brain tissue lysate (male case1) at 20 µg

Lane 3:

Cx3cr1 knockout mouse brain tissue lysate (male case2) 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: 36 kDa

false

Exposure time: 180s