Anti-Heme oxygenase 2/HMOX2 Antibody Picoband®

TLR4 is a negative regulator of biliverdin/BVRA signaling. ( a ) Healthy donor’s blood was untreated (lane 1) or treated with LPS (10 ng/ml) for the indicated times (lanes 2–7). Leukocytes were isolated and analyzed. Abbreviations are: Heme oxygenase 1, HO-1; Heme oxygenase 2, HO-2; Haptoglobin, Hapto. ( b ) In an earlier study , subjects were administered LPS (1 ng/kg) in vivo and blood was drawn at the indicated times post LPS infusion. Leukocyte lysates available from that study were normalized for protein content and analyzed by western blotting. ( c – f ) In another prior study , leukocytes from four subjects administered LPS in vivo were analyzed for changes in gene expression over a period of 24 hours post LPS infusion. Data from that study , available through GEO dataset GSE3284, revealed a temporal ( c ) increase in TLR4 mRNA, ( d ) decline in BVRA mRNA, ( e ) decrease in PKCζ mRNA ( f ) increase in haptoglobin mRNA expression. In ( c – f ) each symbol represents a subject. ( g ) At time 0 (0 hr) healthy donor’s blood was untreated (UN; lane 1), treated for 1 hour with biliverdin (Bili 0 hr; 50 μM; lane 2), treated for 4 hours with LPS (10 ng/ml; lanes 4–6) to trigger a decline in BVRA expression, or for 1 hour with metformin (Met; 10 μM; lane 7). Four hours later (time 4 hr) blood samples were treated for 1 hour with biliverdin (Bili 4 hr; 50 μM; lane 3) or metformin (Met 4 hr; 10 μM; lane 8). Samples pretreated with LPS for 4 hours (lanes 4–6), were then treated for 1 hr with biliverdin (Bili; 50 μM; lane 5) or metformin (Met; 10 μM; lane 6). Leukocytes were isolated and analyzed by western blotting.
Index in PubMed under a CC BY license. PMID: 31065010

Western blot analysis of PHMOX2 using anti-HMOX2 antibody (PB9213).
Electrophoresis was performed on a 10% SDS-PAGE gel at 80V (Stacking gel) / 120V (Resolving gel) for 2 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: huamn Jurkat whole cell lysates,
Lane 2: human Hela whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human PANC-1 whole cell lysates,
Lane 5: rat PC-12 whole cell lysates,
Lane 6: mouse RAW264.7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA 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-HMOX2 antigen affinity purified polyclonal antibody (PB9213) 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 (Catalog # BA1054) at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an ECL Plus Western Blotting Substrate (Catalog # AR1196-200) with Tanon 5200 system. A specific band was detected for HMOX2 at approximately 36 kDa. The expected band size for HMOX2 is at 36 kDa.

IHC analysis of HMOX2 using anti-HMOX2 antibody (PB9213).
HMOX2 was detected in a paraffin-embedded section of human appendix mucinous adenocarcinoma 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-HMOX2 Antibody (PB9213) 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.

IHC analysis of HMOX2 using anti-HMOX2 antibody (PB9213).
HMOX2 was detected in a paraffin-embedded section of human colorectal adenocarcinoma 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-HMOX2 Antibody (PB9213) 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.

IHC analysis of HMOX2 using anti-HMOX2 antibody (PB9213).
HMOX2 was detected in a paraffin-embedded section of human lung adenocarcinoma 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-HMOX2 Antibody (PB9213) 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.

IHC analysis of HMOX2 using anti-HMOX2 antibody (PB9213).
HMOX2 was detected in a paraffin-embedded section of human ovarian serous adenocarcinoma 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-HMOX2 Antibody (PB9213) 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.

IHC analysis of HMOX2 using anti-HMOX2 antibody (PB9213).
HMOX2 was detected in a paraffin-embedded section of human thyroid cancer 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-HMOX2 Antibody (PB9213) 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.