Anti-Hsp60/HSPD1 Antibody Picoband® (monoclonal, 6G2)

Western blot analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
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: human Caco-2 whole cell lysates
Lane 2: human A549 whole cell lysates
Lane 3: human THP-1 whole cell lysates
Lane 4: human SW620 whole cell lysates
Lane 5: human U-937 whole cell lysates
Lane 6: human HepG2 whole cell lysates
Lane 7: rat RH35 whole cell lysates
Lane 8: mouse RAW246.7 whole cell lysates
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 mouse anti-HSPD1 antigen affinity purified monoclonal antibody (Catalog # M01280-3) 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-mouse 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 # EK1001) with Tanon 5200 system. A specific band was detected for HSPD1 at approximately 60KD. The expected band size for HSPD1 is at 60KD.

The differential abundance of HSP90AB1, HSPD1 and HSPA13 in villi tissues was verified by Western blot. A The representative of Western blot analysis to verify selected differentially expressed proteins HSP90AB1, HSPD1 and HSPA13 in the villi tissue of embryo from EMA ( n = 6) and control( n = 6). B The scatter plot of HSP90AB1 abundance in the villi tissue of embryo from EMA ( n = 22) vs control ( n = 22) ( P < 0.01). C The scatter plot of HSPD1 abundance in the villi tissue of embryo from EMA ( n = 22) vs control ( n = 22) ( P < 0.01). D The scatter plot of HSPA13 abundance in the villi tissue of embryo from EMA ( n = 22) vs control ( n = 22) ( P < 0.01)
Index in PubMed under a CC BY license. PMID: 37587463

A , B , C Immunohistochemical staining for HSP90AB1, HSPD1, and HSPA13 observed in the cytoplasm of syncytiotrophoblast and cytotrophoblast cells (× 200, × 400). Villus from 22 patients with EMA showed lower; D - F Quantitative scoring results of IHC analyses w shown as box plots
Index in PubMed under a CC BY license. PMID: 37587463

IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
HSPD1 was detected in paraffin-embedded section of human intestinal cancer tissues. 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 mouse anti-HSPD1 Antibody (M01280-3) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.

IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
HSPD1 was detected in paraffin-embedded section of human lung cancer tissues. 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 mouse anti-HSPD1 Antibody (M01280-3) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.

IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
HSPD1 was detected in paraffin-embedded section of human mammary cancer tissues. 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 mouse anti-HSPD1 Antibody (M01280-3) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.

IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
HSPD1 was detected in paraffin-embedded section of mouse liver tissues. 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 mouse anti-HSPD1 Antibody (M01280-3) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.

IHC analysis of HSPD1 using anti-HSPD1 antibody (M01280-3).
HSPD1 was detected in paraffin-embedded section of rat liver tissues. 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 mouse anti-HSPD1 Antibody (M01280-3) overnight at 4°C. Biotinylated goat anti-mouse 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 # SA1021) with DAB as the chromogen.

Flow Cytometry analysis of A431 cells using anti-HSPD1 antibody (M01280-3).
Overlay histogram showing A431 cells stained with M01280-3 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-HSPD1 Antibody (M01280-3,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Flow Cytometry analysis of HepG2 cells using anti-HSPD1 antibody (M01280-3).
Overlay histogram showing HepG2 cells stained with M01280-3 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-HSPD1 Antibody (M01280-3,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of Hsp60/HSPD1 using anti-Hsp60/HSPD1 antibody (M01280-3).
Hsp60/HSPD1 was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL mouse anti-Hsp60/HSPD1 Antibody (M01280-3) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.