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cd133 antibody a0219  (ABclonal Biotechnology)


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    ABclonal Biotechnology cd133 antibody a0219
    ( a ) FACS plots of <t>CD133</t> and LGR5 staining in patient-derived colorectal cancer stem cells (CRCSCs) expressing the indicated shRNAs and plasmids. See also molecularly characterized CRCSCs derived from patient in . ( b ) Immunofluorescence analysis of the effect of hCINAP knockdown by shRNA on the mesenchymal–epithelial transition in CRCSC spheres. Mesenchymal marker vimentin was labelled in red and epithelial marker E-cadherin was labelled in green. The nucleus was counterstained with DAPI (blue). Scale bar, 100 μm. ( c ) Limiting dilution tumorsphere formation assay in CRCSCs with the indicated shRNAs and plasmids. CRCSC frequency was calculated using extreme limiting dilution analysis. ** P <0.01, *** P <0.001. ( d ) Representative mRNA levels were analysed by qRT–PCR in CRCSCs as indicated. Gene expression in hCINAP-depleted cells was normalized to that of the shRNA control group. The experiments were performed three times and data are determined by Student's t -test and presented as mean±s.e.m. (* P <0.05, ** P <0.01, *** P <0.001, NS, not significant). ( e ) CRCSC spheres were treated with 0.1 μM 5-fluorouracil or 0.1 μM oxaliplatin for 24 h and photographed using a bright field microscope in e . Scale bar, 50 μm. Also see that the viable cell inhibition ratio was determined using MTT colorimetry in .
    Cd133 Antibody A0219, supplied by ABclonal Biotechnology, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cd133+rabbit+polyclonal+a0219+antibody/cd133+antibody/pmc05454382-185-83-88
    Average 90 stars, based on 1 article reviews
    cd133 antibody a0219 - by Bioz Stars, 2026-10
    90/100 stars

    Images

    1) Product Images from "Adenylate kinase hCINAP determines self-renewal of colorectal cancer stem cells by facilitating LDHA phosphorylation"

    Article Title: Adenylate kinase hCINAP determines self-renewal of colorectal cancer stem cells by facilitating LDHA phosphorylation

    Journal: Nature Communications

    doi: 10.1038/ncomms15308

    ( a ) FACS plots of CD133 and LGR5 staining in patient-derived colorectal cancer stem cells (CRCSCs) expressing the indicated shRNAs and plasmids. See also molecularly characterized CRCSCs derived from patient in . ( b ) Immunofluorescence analysis of the effect of hCINAP knockdown by shRNA on the mesenchymal–epithelial transition in CRCSC spheres. Mesenchymal marker vimentin was labelled in red and epithelial marker E-cadherin was labelled in green. The nucleus was counterstained with DAPI (blue). Scale bar, 100 μm. ( c ) Limiting dilution tumorsphere formation assay in CRCSCs with the indicated shRNAs and plasmids. CRCSC frequency was calculated using extreme limiting dilution analysis. ** P <0.01, *** P <0.001. ( d ) Representative mRNA levels were analysed by qRT–PCR in CRCSCs as indicated. Gene expression in hCINAP-depleted cells was normalized to that of the shRNA control group. The experiments were performed three times and data are determined by Student's t -test and presented as mean±s.e.m. (* P <0.05, ** P <0.01, *** P <0.001, NS, not significant). ( e ) CRCSC spheres were treated with 0.1 μM 5-fluorouracil or 0.1 μM oxaliplatin for 24 h and photographed using a bright field microscope in e . Scale bar, 50 μm. Also see that the viable cell inhibition ratio was determined using MTT colorimetry in .
    Figure Legend Snippet: ( a ) FACS plots of CD133 and LGR5 staining in patient-derived colorectal cancer stem cells (CRCSCs) expressing the indicated shRNAs and plasmids. See also molecularly characterized CRCSCs derived from patient in . ( b ) Immunofluorescence analysis of the effect of hCINAP knockdown by shRNA on the mesenchymal–epithelial transition in CRCSC spheres. Mesenchymal marker vimentin was labelled in red and epithelial marker E-cadherin was labelled in green. The nucleus was counterstained with DAPI (blue). Scale bar, 100 μm. ( c ) Limiting dilution tumorsphere formation assay in CRCSCs with the indicated shRNAs and plasmids. CRCSC frequency was calculated using extreme limiting dilution analysis. ** P <0.01, *** P <0.001. ( d ) Representative mRNA levels were analysed by qRT–PCR in CRCSCs as indicated. Gene expression in hCINAP-depleted cells was normalized to that of the shRNA control group. The experiments were performed three times and data are determined by Student's t -test and presented as mean±s.e.m. (* P <0.05, ** P <0.01, *** P <0.001, NS, not significant). ( e ) CRCSC spheres were treated with 0.1 μM 5-fluorouracil or 0.1 μM oxaliplatin for 24 h and photographed using a bright field microscope in e . Scale bar, 50 μm. Also see that the viable cell inhibition ratio was determined using MTT colorimetry in .

    Techniques Used: Staining, Derivative Assay, Expressing, Immunofluorescence, Knockdown, shRNA, Marker, Tube Formation Assay, Quantitative RT-PCR, Gene Expression, Control, Microscopy, Inhibition, Colorimetric Assay

    ( a ) Immunoblot analysis of LDHA Y10 phosphorylation in CRCSC spheres transfected with shRNAs and plasmids as indicated. ( b , c ) Lactate production levels and LDHA enzyme activity were determined in CRCSC spheres as indicated in a . ( d ) ECAR analysis of CRCSCs in a . ( e ) ECAR levels of CRCSCs treated with glucose (10 mM). ( f ) OCR analysis of CRCSCs in a . ( g ) OCR (basal respiration) levels of CRCSCs in a . ( h ) Western blot analysis of LDHA Y10 phosphorylation levels in colon organoid, CRCNSCs (CD133-negative CRC cells derived from patient) and CRCSCs (CD133-positive CRC cells) transfected with scrambled shRNA or hCINAP shRNA, respectively. ( i – m ) ECAR, OCR, NAD + /NADH, ROS and apoptosis levels (quantified from ) were measured from cells in h . Data in b , c , e , g , i – m are are determined by Student's t-test and presented as mean±s.e.m. ( n =3). NS, not significant, * P <0.05, ** P <0.01, *** P <0.001. The unit of ROS (a.u.) is short for arbitrary unit.
    Figure Legend Snippet: ( a ) Immunoblot analysis of LDHA Y10 phosphorylation in CRCSC spheres transfected with shRNAs and plasmids as indicated. ( b , c ) Lactate production levels and LDHA enzyme activity were determined in CRCSC spheres as indicated in a . ( d ) ECAR analysis of CRCSCs in a . ( e ) ECAR levels of CRCSCs treated with glucose (10 mM). ( f ) OCR analysis of CRCSCs in a . ( g ) OCR (basal respiration) levels of CRCSCs in a . ( h ) Western blot analysis of LDHA Y10 phosphorylation levels in colon organoid, CRCNSCs (CD133-negative CRC cells derived from patient) and CRCSCs (CD133-positive CRC cells) transfected with scrambled shRNA or hCINAP shRNA, respectively. ( i – m ) ECAR, OCR, NAD + /NADH, ROS and apoptosis levels (quantified from ) were measured from cells in h . Data in b , c , e , g , i – m are are determined by Student's t-test and presented as mean±s.e.m. ( n =3). NS, not significant, * P <0.05, ** P <0.01, *** P <0.001. The unit of ROS (a.u.) is short for arbitrary unit.

    Techniques Used: Western Blot, Phospho-proteomics, Transfection, Activity Assay, Derivative Assay, shRNA

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    Blocking Assay:

    Article Title: TREM1 is essential for maintaining stemness of liver cancer stem-like cells in hepatocellular carcinoma
    Article Snippet: 80μg of protein were resolved and separated by 4-20% Mini-PROTEAN ® TGXTM Precast gels (45461094, BIO-RAD), transferred onto PVDF membranes (IPFL85R, Millipore). .. Followed by blocking with SuperBlock blocking buffer (PI37580, Thermo Fisher Scientific) for 1h, membranes were incubated with primary antibodies - TREM1 monoclonal (FERMA548755, Invitrogen,1:1000), CD133 Rabbit polyclonal (A0219, Abclonal, 1:1000), Epcam Rabbit polyclonal (A23654, Abclonal, 1:1000), OCT4 Rabbit polyclonal (A7920, Abclonal, 1:1000), Nanog Rabbit polyclonal (A3232, Abclonal, 1:1000), Sox2 Rabbit polyclonal (A19118, Abclonal, 1:1000), β Actin (MAB8929SP, R&D Systems,1:1000), α tubulin (sc-53029, Santa Cruz Biotechnology, 1:1000) at 4°C overnight. .. After washing in TBST, the membranes were incubated with secondary antibodies - goat anti-mouse HRP-conjugated secondary antibody (AP127PMI, Millipore Sigma, 1:10,000), HRP-conjugated Anti-Mouse IgG (MAB8929-SP, R&D Systems, 1:10,000) and HRP-conjugated anti-Rabbit (AS014, Abclonal, 1:10,000) for 1h at room temperature (RT).

    Incubation:

    Article Title: TREM1 is essential for maintaining stemness of liver cancer stem-like cells in hepatocellular carcinoma
    Article Snippet: 80μg of protein were resolved and separated by 4-20% Mini-PROTEAN ® TGXTM Precast gels (45461094, BIO-RAD), transferred onto PVDF membranes (IPFL85R, Millipore). .. Followed by blocking with SuperBlock blocking buffer (PI37580, Thermo Fisher Scientific) for 1h, membranes were incubated with primary antibodies - TREM1 monoclonal (FERMA548755, Invitrogen,1:1000), CD133 Rabbit polyclonal (A0219, Abclonal, 1:1000), Epcam Rabbit polyclonal (A23654, Abclonal, 1:1000), OCT4 Rabbit polyclonal (A7920, Abclonal, 1:1000), Nanog Rabbit polyclonal (A3232, Abclonal, 1:1000), Sox2 Rabbit polyclonal (A19118, Abclonal, 1:1000), β Actin (MAB8929SP, R&D Systems,1:1000), α tubulin (sc-53029, Santa Cruz Biotechnology, 1:1000) at 4°C overnight. .. After washing in TBST, the membranes were incubated with secondary antibodies - goat anti-mouse HRP-conjugated secondary antibody (AP127PMI, Millipore Sigma, 1:10,000), HRP-conjugated Anti-Mouse IgG (MAB8929-SP, R&D Systems, 1:10,000) and HRP-conjugated anti-Rabbit (AS014, Abclonal, 1:10,000) for 1h at room temperature (RT).

    Article Title: Sequential light irradiation-controlled cancer stemness inhibition for sensitized photothermal therapy
    Article Snippet: Cancer stem-like cells (CSCs) have a well-established role in mediating tumor relapse and resistance towards chemotherapy and radiation therapy.. Photothermal therapy (PTT) is an efficient therapeutic strategy that uses light and photothermal agents to generate hyperthermia in tumors and kill cancer cells.. However, due to the heterogeneity and drug resistance of CSCs, some of them may survive from PTT and cause recurrence and metastasis of tumors.

    other:

    Article Title: Adenylate kinase hCINAP determines self-renewal of colorectal cancer stem cells by facilitating LDHA phosphorylation
    Article Snippet: The following antibodies were used in this study: mouse monoclonal Flag antibody (F3165, Sigma, USA; 1:2,000), HA antibody (H9658, Sigma, USA; 1:2,000), mouse monoclonal His antibody (D291-3, MBL, Nagoya, Japan; 1:2,000), mouse monoclonal Myc antibody (M047-3, MBL, Nagoya, Japan; 1:1,000), mouse monoclonal GST antibody (M071-3, MBL, Nagoya, Japan; 1:1,000), rabbit monoclonal ACTIN antibody (PM053, MBL, Nagoya, Japan; 1:2,000), rabbit polyclonal LDHA antibody (BS6179, Bioworld Technology Inc.; 1:2,000), rabbit polyclonal hCINAP antibody (BS2171, Bioworld Technology Inc.; 1:1,000), rabbit polyclonal CD44 antibody (A1351, Abclonal; 1:1,000), rabbit polyclonal CD133 antibody (A0219, Abclonal; 1:1,000), rabbit polyclonal LGR5 antibody (A8090, Abclonal; 1:1,000), rabbit polyclonal HIF1 alpha antibody (GTX127309, GeneTex; 1:500), rabbit polyclonal FGFR1 antibody (I648, Bioworld Technology, Inc.; 1:500) and Goat polyclonal LGR5 antibody (sc-68,580, Santa Cruz Biotechnology; 1:50 for immunofluorescence).



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