Anti-CMKLR1 antibody [EPR26501-70]

• Flow Cyt

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Flow Cytometry - Anti-CMKLR1 antibody [EPR26501-70] (AB306554)

Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) treated with 100ng/ml GM-CSF and 50ng/ml IL-4 for 3 days cells labelling CMKLR1 with ab306554 at 1/50 dilution (1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti-Rabbit IgG (Alexa Fluor^® 647, ab150083) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells. Cells were stained with rabbit IgG or ab306554. Then stained with anti-HLA-DR conjugated to PE.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CMKLR1 antibody [EPR26501-70] (AB306554)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized EA.hy926 (human somatic cell hybrid endothelial) cells labeling CMKLR1 with ab306554 at 1/100 (5.25 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 EA.hy926 cell line ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 ug/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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CMKLR1 antibody [EPR26501-70] (AB306554)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HL-60 (human acute promyelocytic leukemia promyeloblast) cells labeling CMKLR1 with ab306554 at 1/100 (5.25 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 HL-60 cells treated with PMA (50 ng/ml) for 72 h, then add MG-132 (10 uM) for 4 h, and no staining under only MG-132 (10 uM) for 4 h treatment. 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 dilution (2.5 ug/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.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CMKLR1 antibody [EPR26501-70] (AB306554)

Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) treated with 100ng/ml GM-CSF and 50ng/ml IL-4 for 3 days cells labelling CMKLR1 with ab306554 at 1/50 dilution (1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti-Rabbit IgG (Alexa Fluor^® 647, ab150083) at 1/2000 dilution was used as the secondary antibody. Gated on viable HLA-DR and CD123 double positive cells.Cells were stained with rabbit IgG or ab306554. Then stained with anti-CD123 conjugated to FITC and anti-HLA-DR conjugated to PE.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CMKLR1 antibody [EPR26501-70] (AB306554)

Flow cytometric analysis of Human peripheral blood mononuclear cell (PBMC) treated with 100ng/ml GM-CSF and 50ng/ml IL-4 for 3 days cells labelling CMKLR1 with ab306554 at 1/50 dilution (1ug)/ Right compared with a Rabbit monoclonal IgG (ab172730) / Left isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat anti-Rabbit IgG (Alexa Fluor^® 647, ab150083) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells. Cells were stained with rabbit IgG or ab306554. Then stained with anti-CD123 conjugated to FITC.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CMKLR1 antibody [EPR26501-70] (AB306554)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labeling CMKLR1 with ab306554 at 1/100 (5.25 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 weak cytoplasmic staining in Neuro-2a cell line ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor^® 594) was used to counterstain tubulin at 1/200 dilution (2.5 ug/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

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Western blot - Anti-CMKLR1 antibody [EPR26501-70] (AB306554)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Samples are non-boiled as boiling may cause protein aggregation. The expression of Chemokine-like receptor 1 is upregulated in response to PMA treatment (PMID : 19906641). Exposure time : 3 seconds

All lanes:

Western blot - Anti-CMKLR1 antibody [EPR26501-70] (ab306554) at 1/1000 dilution

Lane 1:

HL-60 (human acute promyelocytic leukemia promyeloblast) treated with 10 ?M MG-132 for 4h whole cell lysate 20 µg

Lane 2:

HL-60 treated with /ml PMA for 72h, 10 ?M MG-132 was then added for additional 4h whole cell lysate 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 1/50000 dilution

Observed band size: 42 kDa

false

Exposure time: 3s

• WB

Supplier Data

Western blot - Anti-CMKLR1 antibody [EPR26501-70] (AB306554)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Samples are non-boiled as boiling may cause protein aggregation. Exposure time : 5.5 seconds

All lanes:

Western blot - Anti-CMKLR1 antibody [EPR26501-70] (ab306554) at 1/1000 dilution

Lane 1:

Ea.Hy926 (human somatic cell hybrid endothelial cell) whole cell lysate 20 μg

Lane 2:

HUVEC (human umbilical vein endothelial cell) whole cell lysate 20 μg

Lane 3:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate 20 μg

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate 20 μg

Lane 5:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate 20 μg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 42 kDa

false

Exposure time: 5.5s

• WB

Supplier Data

Western blot - Anti-CMKLR1 antibody [EPR26501-70] (AB306554)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Samples are non-boiled as boiling may cause protein aggregation. Exposure time : 5.5 seconds

All lanes:

Western blot - Anti-CMKLR1 antibody [EPR26501-70] (ab306554) at 1/1000 dilution

Lane 1:

Human tonsil tissue lysate 20 μg

Lane 2:

Human placenta tissue lysate 20 μg

Lane 3:

Mouse spleen tissue lysate 20 μg

Lane 4:

Rat spleen tissue lysate 20 μg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 42 kDa

false

Exposure time: 5.5s

• WB

Supplier Data

Western blot - Anti-CMKLR1 antibody [EPR26501-70] (AB306554)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Samples are non-boiled as boiling may cause protein aggregation. Exposure time : 5.5 seconds

All lanes:

Western blot - Anti-CMKLR1 antibody [EPR26501-70] (ab306554) at 1/1000 dilution

Lane 1:

Human fat tissue lysate 20 μg

Lane 2:

Mouse white fat tissue lysate 20 μg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 42 kDa

false

Exposure time: 5.5s