重组Anti-Cytokeratin 17抗体[EP1623] - Cytoskeleton Marker

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 17 antibody [EP1623] - Cytoskeleton Marker (AB109725)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue using ab109725 at 1/100 dilution

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 17 antibody [EP1623] - Cytoskeleton Marker (AB109725)

Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling Cytokeratin 17 with purified ab109725 at 1/500. Cells were fixed with 100% methanol. ab150077, Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).

Secondary Only Control : PBS was used instead of the primary antibody as the negative control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cytokeratin 17 antibody [EP1623] - Cytoskeleton Marker (AB109725)

Immunohistochemical analysis of paraffin-embedded human squamous cervical carcinoma tissue using ab109725 at 1/100 dilution

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 17 antibody [EP1623] - Cytoskeleton Marker (AB109725)

Immunofluorescent staining of A431 cells using ab109725 at 1/100 dilution

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Cytokeratin 17 antibody [EP1623] - Cytoskeleton Marker (AB109725)

Intracellular Flow Cytometry analysis of A431 (human epidermoid carcinoma) cells labeling Cytokeratin 17 with purified ab109725 at 1/20 dilution (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) (1/2000) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) was used as the unlabeled control.

• WB

Unknown

Western blot - Anti-Cytokeratin 17 antibody [EP1623] - Cytoskeleton Marker (AB109725)

All lanes:

Western blot - Anti-Cytokeratin 17 antibody [EP1623] - Cytoskeleton Marker (ab109725) at 1/1000 dilution

Lane 1:

A431 cell lysate at 10 µg

Lane 2:

HACAT cell lysates at 10 µg

Secondary

All lanes:

Standard HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 48 kDa

false

• ICC/IF

CiteAb

Immunocytochemistry/ Immunofluorescence - Anti-Cytokeratin 17 antibody [EP1623] - Cytoskeleton Marker (AB109725)

Immunocytochemistry-immunofluorescence using Anti-Cytokeratin 17 antibody [EP1623] - Cytoskeleton Marker, ab109725. Publication image from Zhu, L. et al., 2022, Nat Commun, 35440136. Legend direct from paper.

Basal cytokeratins induction is independent of mutant ER genomic binding but requires low ER expression.a Heatmap representing fold change mRNA expression (E2/veh) of six basal cytokeratins and four luminal cytokeratins in ER+ breast cancer lines from six publicly available data sets (GSE89888, GSE94493, GSE108304, GSE3834, GSE38132, and GSE50693). GREB1, PGR, and TFF1 are canonical E2-regulated genes included as positive controls. b Genomic track showing ER binding intensities at KRT5/6A/6B and KRT14/16/17 loci from ER ChIP-seq data sets of MCF7 ESR1 mutant cells. GREB1 locus serve as a positive control. c Graphic view of Pearson correlation between expression of ESR1 and each basal or luminal cytokeratin in ER+ breast tumors in TCGA (n = 808) and METABRIC (n = 1505) cohorts. Color scale and size of dots represent correlation coefficient and significance, respectively. d qRT-PCR measurement of ESR1, KRT5/6A/6B/14/16/17 mRNA levels in MCF7 WT and ESR1 mutant cells with ESR1 siRNA knockdown for 7 days. mRNA fold changes were normalized to WT cells; RPLP0 levels were measured as internal control. Each bar represents mean ± SD with three biological replicates. Data shown are representative from three independent experiments. Student’s t-test (two-sided) was used to compare the gene expression between scramble and knockdown groups. e Representative images of ER, CK5, CK16, and CK17 staining in MCF7-Y537S and D538G cells. BCKs positive cells are highlighted with white arrows. Images were taken under x20 magnification. f Dot plots quantifying the ER intensities in BCKs positive (blue) and the corresponding proximal negative (red) cells from each region. Individual data points from five different regions per group from one experiment, representative of three independent experiments are shown. Paired t-test (two-sided) was applied to compare ER intensities between BCKs positive and negative cells. Source data are provided as a Source Data file for a, c, d, f.