Anti-CCR2 antibody [EPR20844-15]

• Flow Cyt

Lab

Flow Cytometry - Anti-CCR2 antibody [EPR20844-15] (AB273050)

Flow cytometry staining of C57 BL/6 mouse splenocytes with ab273050 (right) or Rabbit monoclonal IgG (ab172730) (left). Cells were incubated for 30 min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab273050 or Rabbit monoclonal IgG (ab172730) (1x 10^6 in 100 μl at 1.0 μg/ml (1/644)) for 30min on ice. The cells were simultaneously stained with Ly6G.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.

• Flow Cyt

Unknown

Flow Cytometry - Anti-CCR2 antibody [EPR20844-15] (AB273050)

Flow cytometric analysis of HEK-293T (human embryonic kidney epithelial cell) (transfected with GFP tagged mouse CCR2 overexpression construct) cells labelling CCR2 with ab273050 at 1/600 dilution (Right) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left). Goat anti rabbit IgG (Alexa Fluor^® 647, ab150079) at 1/2000 dilution was used as the secondary antibody.

Gated on viable cells.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR2 antibody [EPR20844-15] (AB273050)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CCR2 with ab273050 at 1/250 dilution.

Positive staining on mouse spleen

The primary antibody was incubated for 30 mins at room temperature.

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

Counterstained with Hematoxylin.

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR2 antibody [EPR20844-15] (AB273050)

Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling CCR2 with ab273050 at 1/250 dilution.

Positive staining on mouse lung

The primary antibody was incubated for 30 mins at room temperature.

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

Counterstained with Hematoxylin.

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

• Flow Cyt

Lab

Flow Cytometry - Anti-CCR2 antibody [EPR20844-15] (AB273050)

Flow cytometry staining of C57 BL/6 mouse bone marrow cells with ab273050 (right) or Rabbit monoclonal IgG (ab172730) (left). Cells were incubated for 30 min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab273050 or Rabbit monoclonal IgG (ab172730) (1x 10^6 in 100 μl at 0.2 μg/ml (1/3120)) for 30min on ice. The cells were simultaneously stained with Ly6G.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.

• Flow Cyt

Lab

Flow Cytometry - Anti-CCR2 antibody [EPR20844-15] (AB273050)

Flow cytometry staining of C57 BL/6 mouse bone marrow cells with ab273050 (right) or Rabbit monoclonal IgG (ab172730) (left). Cells were incubated for 30 min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab273050 or Rabbit monoclonal IgG (ab172730) (1x 10^6 in 100 μl at 0.2 μg/ml (1/3120)) for 30min on ice. The cells were simultaneously stained with Ly6G.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice.

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.

• Flow Cyt

Lab

Flow Cytometry - Anti-CCR2 antibody [EPR20844-15] (AB273050)

Flow cytometry staining of C57 BL/6 mouse splenocytes with ab273050 (right) or Rabbit monoclonal IgG (ab172730) (left). Cells were incubated for 30 min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab273050 or Rabbit monoclonal IgG (ab172730) (1x 106 in 100 μl at 1.0 μg/ml (1/644)) for 30min on ice. The cells were simultaneously stained with CD11b.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice.

Acquisition of >30000 events were collected using a n/a. Events were gated on viable cells.

• Flow Cyt

Lab

Flow Cytometry - Anti-CCR2 antibody [EPR20844-15] (AB273050)

Flow cytometry staining of C57 BL/6 mouse bone marrow cells with ab273050 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were incubated for 30 min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab273050 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 μl at 0.2 μg/ml (1/12000)) for 30min on ice. The cells were simultaneously stained with CD11b.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR2 antibody [EPR20844-15] (AB273050)

Immunohistochemical analysis of paraffin-embedded (A) Lymph node tissue from wild-type C57BL/6JGpt mice (B) Lymph node tissue from CCR2 knockout mice staining with ab273050 at 1/300 dilution and ready-to-use Goat Anti-Rabbit IgG H&L (HRP) secondary. Counterstaining with hematoxylin. Heat mediated antigen retrieval was performed with Citrate buffer (pH 6.0, Epitope Retrieval Solution 1) for 20 mins. Positive staining on (A) Lymph node tissue from wild-type C57BL/6JGpt mice and no staining on (B) Lymph node tissue from CCR2 knockout mice. The section was incubated with ab273050 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND™ RX instrument. The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and CCR2-KO homozygous mice (Strain ID : T006112).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CCR2 antibody [EPR20844-15] (AB273050)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CCR2 with ab273050 at 1/250 dilution.

Positive staining on rat spleen

The primary antibody was incubated for 30 mins at room temperature.

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

Counterstained with Hematoxylin.

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

• IP

Unknown

Immunoprecipitation - Anti-CCR2 antibody [EPR20844-15] (AB273050)

CCR2 was immunoprecipitated from 0.35 mg Mouse lymph node tissue lysate with ab273050 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab273050 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/1000 dilution.

Lane 1 : Mouse lymph node tissue lysate 10 ug.

Lane 2 : ab273050 IP in Mouse lymph node tissue lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab273050 in mouse lymph node tissue lysate.

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

Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-CCR2 antibody [EPR20844-15] (ab273050)

Predicted band size: 42 kDa

Observed band size: 40 kDa

false

• WB

Lab

Western blot - Anti-CCR2 antibody [EPR20844-15] (AB273050)

The tissue samples were kindly provided by GemPharmatech. C57BL/6JGpt wildtype mice and CCR2-KO homozygous mice (Strain ID : T006112).

All lanes:

Western blot - Anti-CCR2 antibody [EPR20844-15] (ab273050) at 1/1000 dilution

Lane 1:

Wild-type mouse spleen tissue lysate (male)

Lane 2:

CCR2 knockout mouse spleen tissue lysate (male)

Lane 3:

Wild-type mouse lung tissue lysate (male)

Lane 4:

CCR2 knockout mouse lung tissue lysate (male)

Lane 5:

Wild-type mouse spleen tissue lysate (female)

Lane 6:

CCR2 knockout mouse spleen tissue lysate (female)

Lane 7:

Wild-type mouse lung tissue lysate (female)

Lane 8:

CCR2 knockout mouse lung tissue lysate (female)

false

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CCR2 antibody [EPR20844-15] (AB273050)

Immunofluorescent analysis of 4% Paraformaldehyde fixed 0.1% Triton X-100 permeabilized 293T (Human embryonic kidney epithelial cell) transfected with GFP tagged mouse CCR2 overexpression construct labelling CCR2 with ab273050 at 1/250 dilution followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) as secondary at 1/1000 (2 μg/ml) dilution. Confocal image showing membranous and cytoplasmic staining in HEK-293T cells transfected with with GFP tagged mouse CCR2 overexpression construct (shown in Red). GFP signals shown in Green. Nuclear DNA was labelled with DAPI (shown in blue).