Anti-Cytokeratin 18/KRT18 Antibody Picoband®

Characterization of hAECs, detection of the survival hAECs encapsulated in SA-BG, and schematic illustration of the surgical procedure. a The morphology of cultured hAECs was observed under a microscope. Scale bar 100 μm. b Flow cytometry analysis of cell surface markers on hAECs. The isotypes (ISO) of SSEA4 and CD324 were used as negative controls. c Immunostaining images showed the high expression of epithelial marker (CK18) and stem cell marker (TRA-1-60). Scale bar 100 μm. d The fabrication method of SA-BG-loaded hAECs and CM. e Representative live/dead images of hAECs encapsulated in SA-BG at days 1, 7, 10, and 14, respectively. hAECs encapsulated in SA-BG treated with 70% alcohol were positive for PI. Live cells were shown green color and dead cells were red color. Scale bar 100 μm. f Bright field image of hAECs capsulated in SA-BG at day 1 ( a ) and 14 ( b ). Scale bar 100 μm. g The percentage of live cells to total cells. h Schematic of the experimental procedure for the transplantation of SA-BG-loaded hAECs/CM into mice with chemotherapy-induced POF
Index in PubMed under a CC BY license. PMID: 33794993

The effect of SA-BG extracts on the biological characterization and paracrine capacity of hAECs. a The viability of hAECs cultured with SA-BG extracts was detected by CCK-8 assay. b – d Expression of stemness (Oct-4 and Nanog) and epithelial (CK18) genes of hAECs cultured with SA-BG extracts at different time points. e , f The results of cytokine array of CM from hAECs and SA-BG extract-treated hAECs, respectively. g Column displayed the higher expression of cytokines in the SA-BG extract-treated hAECs than in the control hAECs. h , i The results of quantification of angiogenic factors released from SA-BG extract-treated hAECs and control hAECs by ELISA
Index in PubMed under a CC BY license. PMID: 33794993

hAECs express specific surface markers and have stem cell characteristics with low immunogenicity. a hAECs presented an epithelial morphology under bright-field microscopy. Scale bar = 100 μm. b hAECs were labeled with CFSE to track implanted cells. The expression rate of green fluorescence staining was nearly 100%. Scale bar = 100 μm. c-f By flow cytometry, hAECs were positive for stem cell marker SSEA-4 ( c ) and epithelial marker CD324 ( d ) and were negative for mesenchymal markers CD146 ( e ) and HLA-DR ( f ). g Immunofluorescence staining for CK18 (an epithelial marker) expression and vimentin (a mesenchymal marker) in hAECs. Scale bar = 200 μm
Index in PubMed under a CC BY license. PMID: 33762002

Western blot analysis of KRT18 using anti-KRT18 antibody (A01357-1).
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: human placenta tissue lysates,
Lane 2: human CACO2 whole cell lysates,
Lane 3: human A549 whole cell lysates,
Lane 4: human A431 whole cell lysates,
Lane 5: rat liver tissue lysates,
Lane 6: rat thymus tissue lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse thymus tissue 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-KRT18 antigen affinity purified polyclonal antibody (A01357-1) 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 KRT18 at approximately 48 kDa. The expected band size for KRT18 is at 48 kDa.

IHC analysis of KRT18 using anti-KRT18 antibody (A01357-1).
KRT18 was detected in a paraffin-embedded section of human intestinal 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-KRT18 Antibody (A01357-1) 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 KRT18 using anti-KRT18 antibody (A01357-1).
KRT18 was detected in a paraffin-embedded section of human intestinal 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-KRT18 Antibody (A01357-1) 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 KRT18 using anti-KRT18 antibody (A01357-1).
KRT18 was detected in a paraffin-embedded section of human breast 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-KRT18 Antibody (A01357-1) 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 KRT18 using anti-KRT18 antibody (A01357-1).
KRT18 was detected in a paraffin-embedded section of human placenta 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-KRT18 Antibody (A01357-1) 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 KRT18 using anti-KRT18 antibody (A01357-1).
KRT18 was detected in a paraffin-embedded section of mouse intestine 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-KRT18 Antibody (A01357-1) 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 KRT18 using anti-KRT18 antibody (A01357-1).
KRT18 was detected in a paraffin-embedded section of rat colon 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-KRT18 Antibody (A01357-1) 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.

IF analysis of KRT18 using anti-KRT18 antibody (A01357-1)
KRT18 was detected in paraffin-embedded section of human intestinal cancer tissues. 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 5μg/mL rabbit anti-KRT18 Antibody (A01357-1) overnight at 4°C. Biotin conjugated goat anti-rabbit IgG (BA1003) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Cy3 Conjugated Avidin (BA1037). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of KRT18 using anti-KRT18 antibody (A01357-1)
KRT18 was detected in paraffin-embedded section of human gastric cancer tissues. 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 5μg/mL rabbit anti-KRT18 Antibody (A01357-1) overnight at 4°C. Biotin conjugated goat anti-rabbit IgG (BA1003) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Cy3 Conjugated Avidin (BA1037). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of KRT18 using anti-KRT18 antibody (A01357-1)
KRT18 was detected in paraffin-embedded section of human lung cancer tissues. 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 5μg/mL rabbit anti-KRT18 Antibody (A01357-1) overnight at 4°C. Biotin conjugated goat anti-rabbit IgG (BA1003) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Cy3 Conjugated Avidin (BA1037). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of KRT18 using anti-KRT18 antibody (A01357-1)
KRT18 was detected in paraffin-embedded section of human prostatic cancer tissues. 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 5μg/mL rabbit anti-KRT18 Antibody (A01357-1) overnight at 4°C. Biotin conjugated goat anti-rabbit IgG (BA1003) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Cy3 Conjugated Avidin (BA1037). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of KRT18 using anti-KRT18 antibody (A01357-1)
KRT18 was detected in paraffin-embedded section of human ovarian cancer tissues. 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 5μg/mL rabbit anti-KRT18 Antibody (A01357-1) overnight at 4°C. Biotin conjugated goat anti-rabbit IgG (BA1003) was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Cy3 Conjugated Avidin (BA1037). The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow Cytometry analysis of A431 cells using anti-KRT18 antibody (A01357-1).
Overlay histogram showing A431 cells stained with A01357-1 (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 rabbit anti-KRT18 Antibody (A01357-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. Fluoro488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.