Anti-CD13 antibody [EPR27472-288]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD13 antibody [EPR27472-288] (AB317440)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) cells labelling CD13 with ab317440 at 1/50 (10.06 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 staining in J774A.1 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Negative control : EL4 (PMID : 8103749).
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 2.5 ug/ml dilution (Magenta). 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-CD13 antibody [EPR27472-288] (AB317440)

Flow cytometric analysis of Mouse spleen cell cells labelling CD13 with ab317440 at 1/500 dilution (0.1ug) / 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® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with anti-CD3 conjugated to Alexa Fluor®647.
Gated on viable cells.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD13 antibody [EPR27472-288] (AB317440)

Flow cytometric analysis of Mouse spleen cell cells labelling CD13 with ab317440 at 1/500 dilution (0.1ug) / 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® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Cells were co-stained with anti-CD11c conjugated to Brilliant Violet 421.
Gated on viable cells.

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD13 antibody [EPR27472-288] (AB317440)

Flow cytometric analysis of J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage) (Right) / EL4 (mouse lymphoma T lymphocyte) (Left) cells labelling CD13 with ab317440 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cells.
Negative control : EL4 (PMID : 8103749)

• IP

Supplier Data

Immunoprecipitation - Anti-CD13 antibody [EPR27472-288] (AB317440)

CD13 was immunoprecipitated from 0.35 mg J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage ) whole cell lysate with ab317440 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317440 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage ) whole cell lysate
Lane 2 : ab317440 IP in J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage ) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab317440 in J774A.1 whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-CD13 antibody [EPR27472-288] (ab317440) at 1/30 dilution

All lanes:

J774A.1 (mouse reticulum cell sarcoma monocyte/macrophage ) 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: 24s

• WB

Supplier Data

Western blot - Anti-CD13 antibody [EPR27472-288] (AB317440)

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

Negative control : EL4 (PMID : 8103749).

Low expression : heart.

The molecular weight observed is consistent with what has been described in the literature (PMID : 37942010, 8103749).

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

All lanes:

Western blot - Anti-CD13 antibody [EPR27472-288] (ab317440) at 1/1000 dilution

Lane 1:

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

Lane 2:

EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Mouse heart 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

Observed band size: 150 kDa,36 kDa

false

Exposure time: 81s