Anti-Lipocalin-2 / NGAL antibody [EPR5084]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (AB125075)

Immunocytochemistry analysis of SW480 (human colorectal adenocarcinoma epithelial cell) labeling Lipocalin-2 / NGAL with purified ab125075 at 1/250 dilution (10 µg/ml). Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/1000 (2 µg/ml) was used as the secondary antibody. ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.9 µg/ml) was used as counterstain. Nuclei were stained blue with DAPI.
Negative control : PBS instead of the primary antibody.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (AB125075)

Immunohistochemical analysis of paraffin-embedded human kidney tissue sections labeling Lipocalin-2 / NGAL with purified ab125075 at 1/100 dilution. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Sections were counterstained with Hematoxylin.
Antigen retrieval was heat mediated antigen retrieval using citrate buffer, pH 6.0).

• WB

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Western blot - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (AB125075)

All lanes:

Western blot - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075) at 1/1000 dilution

Lane 1:

Human fetal kidney lysate at 10 µg

Lane 2:

SW480 (Human colorectal adenocarcinoma cell line) cell lysate at 10 µg

Predicted band size: 22 kDa

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• WB

Lab

Western blot - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (AB125075)

Lanes 1 - 4 : Merged signal (red and green). Green - ab125075 observed at 23 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab125075 was shown to react with Lipocalin-2 / NGAL in wild-type SW480 cells in Western blot with loss of signal observed in LCN2 knockout cell line ab270486 (knockout cell lysate ab270509). Wild-type SW480 and LCN2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab125075 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Lipocalin-2 / NGAL antibody [EPR5084] (ab125075) at 1/1000 dilution

Lane 1:

Wild-type SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

LCN2 knockout SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human LCN2 knockout SW480 cell line (<a href='/en-us/products/cell-lines/human-lcn2-knockout-sw480-cell-line-ab270486'>ab270486</a>)

Lane 3:

A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

SH-SY5Y (Human neuroblastoma cell line from bone marrow) whole cell lysate at 20 µg

Predicted band size: 22 kDa

Observed band size: 23 kDa

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