Anti-Lipocalin 2/LCN2 Antibody Picoband®

Western blot analysis of Lipocalin 2 using anti-Lipocalin 2 antibody (PB9609).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50ug of sample under reducing conditions.
Lane 1: Mouse Lung Tissue Lysate,
Lane 2: Mouse Intestine Tissue Lysate.
After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Lipocalin 2 antigen affinity purified polyclonal antibody (Catalog # PB9609) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Lipocalin 2 at approximately 22KD. The expected band size for Lipocalin 2 is at 22KD.

IHC analysis of Lipocalin 2 using anti-Lipocalin 2 antibody (PB9609).
Lipocalin 2 was detected in paraffin-embedded section of Mouse Spleen Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Lipocalin 2 Antibody (PB9609) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

IHC analysis of Lipocalin 2 using anti-Lipocalin 2 antibody (PB9609).
Lipocalin 2 was detected in paraffin-embedded section of Rat Spleen Tissue. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Lipocalin 2 Antibody (PB9609) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

IHC analysis of Lipocalin 2 using anti-Lipocalin 2 antibody (PB9609).
Lipocalin 2 was detected in frozen section of mouse lung tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Lipocalin 2 Antibody (PB9609) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

IHC analysis of Lipocalin 2 using anti-Lipocalin 2 antibody (PB9609).
Lipocalin 2 was detected in frozen section of mouse spleen tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Lipocalin 2 Antibody (PB9609) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

IHC analysis of Lipocalin 2 using anti-Lipocalin 2 antibody (PB9609).
Lipocalin 2 was detected in frozen section of rat lung tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Lipocalin 2 Antibody (PB9609) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

IHC analysis of Lipocalin 2 using anti-Lipocalin 2 antibody (PB9609).
Lipocalin 2 was detected in frozen section of rat spleen tissue. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1μg/ml rabbit anti-Lipocalin 2 Antibody (PB9609) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

IHC analysis of Lipocalin 2 using anti-Lipocalin 2 antibody (PB9609).
Lipocalin 2 was detected in a paraffin-embedded section of mouse kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Lipocalin 2 Antibody (PB9609) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Effects of Nrf2 on MCTR1-regulated ferroptosis in CLP-induced AKI. Mice were given ML385 (30 mg/kg, ip, qd) for 7 d. On day 7, ML385 injection with or without a twice-daily administration mode of MCTR1 as described before in the following experiment. All kidney samples were collected at 24 h after CLP. A–D Shown are representative western blotting and quantification of Nrf2, GPX4, and PTGS2. E – G Quantitative analyses of MDA, GSH, and non-heme iron. H Quantitative analyses of Perl’s stain. I Representative Perl’s stain images. J Representative TEM images. The black arrow indicates ferroptosis-like mitochondria. K Quantitative analyses of mitochondrial length. L Histological analyses of renal tubular injury. M , N Representative HE stains and PAS stain. O Representative IHC images for NGAL. P Quantitative analyses of IHC stain of NGAL. Q , R Quantitative analyses of serum Scr and BUN. n = 6 mice/group, mean ± SD were presented. **P < 0.01
Index in PubMed under a CC BY license. PMID: 34961563

Effects of MCTR1 on ferroptosis in CLP-induced AKI. Mice were given MCTR1 once, twice, or not during the 24 h experiment. The once-daily administration mode (qd): Mice were given MCTR1 (200 ng/mice, iv) 0.5 h before being subjected to CLP. The twice-daily administration mode (bid): Mice were given MCTR1 (200 ng/mice, iv) 0.5 h before being subjected to CLP, and then an additional MCTR1 (200 ng/mice, iv) was given 12 h after CLP. All kidney samples were collected at 24 h after CLP. A , B Shown are representative western blotting and quantification of GPX4 and PTGS2. C–E Quantitative analyses of MDA, GSH, and non-heme iron. F Quantitative analyses of Perl’s stain. G Representative Perl’s stain images. H Representative TEM images. The black arrow indicates ferroptosis-like mitochondria. I Quantitative analyses of mitochondrial length. J Histological analyses of renal tubular injury. K , L Shown are representative HE stains and PAS stain. M Representative IHC images for NGAL. N Quantitative analyses of IHC stain of NGAL. O , P Quantitative analyses of serum Scr and BUN. n = 6 mice/group, mean ± SD were presented. **P < 0.01 . iv: intravenous
Index in PubMed under a CC BY license. PMID: 34961563

Ferroptosis is present in CLP-induced AKI. Mice were subjected to CLP, then the kidneys were collected at the indicated time. A , B Shown are representative western blotting and quantification of GPX4 and PTGS2. C–E Quantitative analyses of MDA, GSH, and non-heme iron. F Quantitative analyses of Perl’s stain. G Representative Perl’s stain images. H Representative transmission electron microscopy (TEM) images. The black arrow indicates ferroptosis-like mitochondria. I Quantitative analyses of mitochondrial length. Mice were pretreated with liproxstatin-1 (10 mg/kg, ip) or DFO (20 mg/kg, ip) 0.5 h before being subjected to CLP. All kidney samples were collected at 24 h after CLP. J–K Shown are representative Hematoxylin-eosin (HE) stains and Periodic Acid-Schiff (PAS) stain. L Histological analyses of renal tubular injury. M Quantitative analyses of IHC stain of NGAL. N Representative immunohistochemistry (IHC) images for NGAL. O , P Quantitative analyses of Scr and BUN. n = 6 mice/group, mean ± SD were presented. *P < 0.05, **P < 0.01 and ns : P > 0.05 . ip: intraperitoneal
Index in PubMed under a CC BY license. PMID: 34961563