11-0247-42

antibody from Invitrogen Antibodies

Targeting: CD24 CD24A

Provider product page for 11-0247-42

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Antibody data

Antibody Data
Antigen structure
References [27]
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Validations
Flow cytometry [2]
Other assay [28]

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Product number: 11-0247-42 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD24 Monoclonal Antibody (eBioSN3 (SN3 A5-2H10)), FITC, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The SN3 monoclonal antibody reacts with the human CD24 molecule, also known as Heat Stable Antigen (HSA). This 35-50 kDa molecule is anchored in the plasma membrane via phosphatidylinositol and is expressed by erythrocytes, thymocytes, peripheral lymphocytes and myeloid lineage. It has been reported that P-selectin (CD62P) binds to CD24. Applications Reported: The SN3 antibody has been reported for use in flow cytometric analysis. Applications Tested: This eBioSN3 (SN3 A5-2H10) antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Excitation: 488 nm; Emission: 520 nm; Laser: Blue Laser. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human
Host: Mouse
Conjugate: Green dye
Isotype: IgG
Antibody clone number: eBioSN3 (SN3 A5-2H10)
Vial size: 100 Tests
Concentration: 5 μL/Test
Storage: 4°C, store in dark, DO NOT FREEZE!

CD24 protein structure - 11-0247-42 shown in red.

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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: NULL
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 2 GPT2-induced breast cancer cell stemness and tumorigenesis A. & B. GPT2 promotes colony formation of breast cancer cells. A. Colony formation assay (n = 3), ***: P < 0.001; **: P < 0.01. B. Soft agar assay (n = 3). ***: P < 0.001 C. The effects of glutamine on colony formation. The basic medium was without glutamine. D. Flow cytometry analysis of CD44 and CD24 expression in GPT2 overexpressing MDA-MB-231 and GPT2 knockdown MCF7 cells. The values represent percentage of CD44 + CD24 - cells. **: P < 0.01;*: P < 0.05. E. GPT2 and ALDH expression was analyzed in GPT2 overexpressing MDA-MB-231 and GPT2 knockdown MCF7 cells by immunofluorescence staining. Red and green represent GPT2 and ALDH, respectively (20 X). F. Representative Western blots show the expression of ALDH, SOX2, OCT4 and NANOG in GPT2 overexpressing MDA-MB-231 and GPT2 knockdown MCF7 cells. G. GPT2 and ALDH expression was analyzed in clinical breast tumors and paratumor tissues by immunofluorescence staining (n= 6 cases). Red and green represent GPT2 and ALDH, respectively (10 X). The values on the histogram represent cell numbers per view. The big white bars represent GPT2 positive cell numbers while the small shading bars inside indicate ALDH positive cell numbers. GPT2 overexpression cells were MDA-MB-231 while GPT2 knockdown cells were MCF7. Data in (A-G) represent three independent experiments. Error bars: standard deviation. ***: P < 0.001; **: P < 0.01; *: P < 0.05.
Conjugate: Green dye

Supportive validation

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Experimental details: Figure 4 obASCs promote a cancer stem-like phenotype through leptin. ( A ) Transwell co-culture with shCtrl obASCs upregulates NOTCH1, NOTCH3, DELTA1, and JAGGED2 in ER + BCCs. ( B ) BCCs have increased ability to form mammospheres after co-culture with shCtrl obASCs followed by 2 Gy radiation compared to non-co-cultured cells or cells co-cultured with shLep obASCs. ( C ) Flow cytometric analysis of the breast cancer stem cell markers CD44 + CD24 - was evaluated for BCCs after transwell co-culture with shCtrl obASCs or shLep obASCs. Values reported are the mean of three independent experiments each performed in triplicate. Bars, +- SEM. * p < 0.05, ** p < 0.01, *** p < 0.001.
Conjugate: Green dye

Supportive validation

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Experimental details: Figure 2 CD10 in CAFs sustains cancer stemness and chemoresistance. A-E) Indicated tumor cell lines were cultured alone (-) or cocultured with CD10 + GPR77 + -depleted CAFs (CD10 + GPR77 + -d) or paired CD10 + GPR77 + CAFs transduced without (-) or with shGFP or shCD10. A) Representative images of mammosphere formation in MCF-7 cells. Scale bar, 100 um. B) Representative images of PKH26 and Numb immunofluorescence staining of MCF-7 cells. Scale bar, 50 um. C,D) Percentage of ALDH1 + cells in C) MCF-7 and D) BT-549 cells detected by flow cytometry. Numerical values are presented as percentage (mean +- SEM, n = 3). E) Percentage of CD44 high CD24 low cells in MCF-7 cells cocultured with indicated CAFs was detected by flow cytometry. Numerical values are presented as percentage (mean +- SEM, n = 3). F,G) Representative flow cytometry plots for F) MCF-7 and G) BT-549 cells treated with docetaxel after being cultured alone (-) or cocultured with the indicated CAFs. The proportion of Annexin V + /Propidium iodide - (early apoptosis) and Annexin V + /Propidium iodide + (late apoptosis) cells is shown. The numerical values indicate Annexin V + percentage. Data are represented as the mean +- SEM of F) n = 4 or G) n = 3 independent experiments. H) Representative immunoblots for cleaved/total caspase-3 and PARP in SK-BR3 cells treated with docetaxel after being cultured alone (-) or cocultured with indicated CAFs ( n = 3). I-L) MCF-7 cells were cocultured with CD10 + GPR77 + CAFs or CD1
Conjugate: Green dye

Supportive validation

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Experimental details: Fig. 2 Cancer stem cell (CSC)-like cells are enriched in the hypoxic populations freshly isolated from xenografts. Tumor cells are enzymatically dissociated and isolated from either the MDA-MB-231/HRE-EGFP ( a - c ) or MCF7/HRE-EGFP ( d - f ) xenografts. Stem cell characteristics are evaluated by fluorescence-activated cell sorting (FACS) for the expression of CSC-associated surface markers CD24, CD44 and CD49f. Representative FACS plots are shown in a , c , d and f . Quantitative population analyses are shown in b ( n = 4-5; * p < 0.05, *** p < 0.001, Student's t test) and e (n = 4; *** p < 0.001, Student's t test). These results are confirmed by three or more independent experiments. EGFP, enhanced green fluorescent protein; SSC, side scatter
Conjugate: Green dye

Supportive validation

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Experimental details: Fig. 4 The cancer stem cell (CSC)-like population is further enriched in secondary xenografts derived from the enhanced green fluorescent protein (EGFP) + MDA-MB-231 cells. a Generation of the secondary xenografts. b , c Surface levels of CD24 and CD44 are analyzed by fluorescence-activated cell sorting. b Average CD24 + populations from six individual tumors (*** p < 0.001, Student's t test). c Average CD44 ++ (right-pointing arrow) populations from three individual tumors (* p < 0.05, Student's t test). d Quantitative RT-PCR analysis of expression of CD24 and CD44 genes in the EGFP + and EGFP - cells freshly isolated from either the 2 nd or 1 st xenografts (n = 3; * p < 0.05, ** p < 0.01, Student's t test). e Clonogenic growth of sorted EGFP + and EGFP - cells freshly isolated from the 2 nd xenografts in comparison to the unsorted tumor cells from the 1 st xenografts (n = 3; * p < 0.05, *** p < 0.0001, Student's t test)
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 5 The cancer stem cell (CSC)-like characteristics of tumor cells isolated from the secondary enhanced green fluorescent protein (EGFP) + MCF7/HRE-EGFP xenografts. a Generation of secondary MCF7/HRE-EGFP xenografts by re-implantation of sorted EGFP + and EGFP - cells isolated from the primary MCF7/HRE-EGFP xenografts. Tumor cells freshly isolated from the secondary xenografts were sorted into EGFP + and EGFP - populations for ( b ) fluorescence-activated cell sorting (FACS) analysis of the CD44 + /CD24 + and CD44 + populations ( n = 5 for EGFP + cells, n = 4 for EGFP - cells; *** p < 0.001, Student's t test). c Side population (SP) of the secondary xenograft-derived tumor cells. MCF7 tumor cells were isolated from the secondary xenografts derived from EGFP + and EGFP - tumor cells, respectively, and expanded in vitro for three passages. Cells were stained with Hoechst 33342 for side population analysis by FACS. Verapamil (50 muM) was used to block nuclear export of Hoechst 33342. These results were validated in two independent experiments. d Clonogenic potential of the freshly sorted EGFP + and EGFP - populations from the secondary xenografts ( n = 6; **** p < 0.0001, *** p < 0.001, Student's t test)
Conjugate: Green dye

Supportive validation

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Experimental details: Figure 4 Down-regulation of BRCA1 promotes cancer cell stemness in neuroblastoma cells. BRCA1 was down-regulated by RNA inference in the SK-N-BE(2)C human neuroblastoma cells (Western blot shown in A ). Effects of BRCA1 on neuroblastoma cell clonogenicity was measured by the clonogenic assay ( B , n = 6). The cell surface levels of CD44 ( C ) and CD24 ( D ) in BE(2)C +- siBRCA1 cells were analyzed by flow cytometry (n = 3). Expression of neuroendocrine cancer stem cell-associated genes in BE(2)C +- siBRCA1 cells were measured by qRT-PCR ( E , n = 3).
Conjugate: Green dye

Supportive validation

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Experimental details: Fig. 4 IGF2BP2-mediated upregulation of DANCR promotes cell proliferation and stemness-like properties. a IGF2BP2 siRNA down regulates DANCR expression in BXPC-3 cells, as determined by qRT-PCR, whereas ectopic expression of IGF2BP2 increases DANCR level ( b ). c Like IGF2BP2 siRNA, IGF2BP2 KO also suppresses DANCR expression in BXPC-3 cells. d DANCR promotes cell proliferation in BXPC-3 and SW1990 cells, as determined by CCK8 assays. e DANCR also promotes colony formation. f Ectopic expression of DANCR increases sphere formation in number and size. g DANCR-overexpression promotes expression of stemness-related genes (OCT4, NANOG, BMI 1, CD24, and CD133) in pancreatic cancer cells, as determined by qRT-PCR and at the protein level ( h ). i Percentage of CD24 + and CD133 + population in DANCR overexpression and control (NC) cells, as determined by flow cytometry. The statistical results of flow cytometry were showed on the right. Values in a , b , c , d , e , g , i are mean +- SEM. * P < 0.05; ** P < 0.01; *** P < 0.001.
Conjugate: Green dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 5 DANCR KO suppresses cell proliferation and stemness-like properties of pancreatic cancer cells. a DANCR KO suppresses cell proliferation in BXPC-3 and SW1990 cells, as determined by CCK8 assays. b DANCR KO suppresses colony formation. c DANCR KO inhibits sphere formation in number and size. d DANCR KO suppresses stemness-related genes in pancreatic cancer cells, as determined by qRT-PCR and at the protein level e . f Percentage of CD24 + and CD133 + population in DANCR KO and control (NC) cells, as determined by flow cytometry. The statistical results of flow cytometry were showed on the right. Values in a , b , d , e , f are mean +- SEM. * P < 0.05; ** P < 0.01; *** P < 0.001.
Conjugate: Green dye

Supportive validation

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Experimental details: Figure 3 Invasion assay of these four subsets of MCF7 cells. A , Cell number quantification of invasion. B , Confocal images of these four subsets of MCF7 cells (Green: CD44 + /CD24 -/low ; Red: CD44 - /CD24 + ; Yellow: CD44 + /CD24 + ; and Blue: CD44 - /CD24 - ). C , Quantification of tumor cell migration distance. D , Images of cell migration distance.
Conjugate: Green dye

Supportive validation

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Experimental details: Effect of naringin on TNF- alpha -induced human NP cells. (a) Chemical structure of naringin. (b) Identification of human NP cell marker CD24 by flow cytometry. (c) CCK-8 results of human NP cells treated with different concentrations of naringin contain 0.1% DMSO for 24 h. (d) CCK-8 results of naringin pretreated human NP cells induced by TNF- alpha for 24 h. The data represent the means +- SD. Significant differences between groups are indicated as * p < 0.05, ** p < 0.01, and *** p < 0.001, n = 3.
Conjugate: Green dye

Supportive validation

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Experimental details: Fig. 4 B3GNT5 promoted mammosphere formation and enhances CSC population. A , B Mammosphere formation of BT549 and MDA-MB231 cells with or without B3GNT5 knockout as well as stably B3GNT5-expressing KO cells ( A ), and representative images are shown ( B ). Scale bars, 50 mum. ( C ) Populations of CSCs (CD44 high /CD24 low ) were measured by flow cytometry in MDA-MB231 and BT549 cells with or without B3GNT5 knockout as well as stably B3GNT5-expressing KO cells. D Expression of B3GNT5 was analyzed by Western blotting in MCF7 cells with stable empty vector or B3GNT5 expression. E , F Mammosphere formation ( E ) and population of CSCs (CD44 high /CD24 low ) ( F ) were examined in MCF7 cells with stable empty vector or B3GNT5-WT-Flag cDNA. G Sequential mammosphere formation were measured in MDA-MB231 and BT549 cells with or without B3GNT5 knockout as well as stably B3GNT5-expressing KO cells. P1, P2 and P3 indicated primary, secondary and tertiary passage of mammosphere, respectively. H Limiting dilution sphere forming assay was performed by fluorescence-activated cell sorting in MDA-MB231 KO and BT549 KO cells with B3GNT5-WT-mcherry overexpression. Cells with high (mcherry high ) and low (mcherry low ) B3GNT5 expression were sorted separately. Data are presented as the percentage of wild-type or vector cell lines. * p < 0.05 by Student's t test. Data are shown as mean +- SD based on three independent experiments
Conjugate: Green dye

Supportive validation

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Experimental details: Figure 3 Stable knockdown of TFAP2C induced EMT MCF-7 cell clone 45 derived by stable expression of shRNA targeting TFAP2C (sKD-C) compared to cell clone derived with non-targeting shRNA (sKD-NT). The clone used for all data in this figure was sKD-C-clone 45. A. Expression pattern of TFAP2A and TFAP2C by RT-PCR in stable cell clones. B. Cell morphology with using the brightfield microscopy 400x magnification. C. Section of expression array comparing sKD-NT vs. sKD-C cells demonstrates down-regulation of Luminal Differentiation Markers and up-regulation of Mesenchymal Markers. D. Western blot showing protein expression of AP-2 proteins and examples of luminal differentiation and mesenchymal markers. E. Relative expression of CD44 gene by RT-PCR for TFAP2C shows increased CD44 RNA in sKD-C cells, confirming array in A and protein in D. F . Distribution of CD44/CD24 subpopulations in stable cell clones of MCF7 by flow cytometry demonstrate an increase in the CD44 +/hi /CD24 -/low population.
Conjugate: Green dye

Supportive validation

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Experimental details: Figure 2 Effects of SATB1 expression on the CD44 + /CD24 - population and invasive properties of human breast cancer cell lines. Populations of CD44 + /CD24 - in (A) MCF-7 cells with SATB1 overexpression and (B) BT-549 cells with SATB1 knockdown, with representative images from the CD44/CD24 flow cytometric analysis. Invasiveness in (C) MCF-7 cells with SATB1 overexpression and (D) BT-549 cells with SATB1 knockdown, and representative images taken from the invasion assays (magnification, x20). All data are presented as the mean +- standard deviation of two independent experiments in triplicate; * P
Conjugate: Green dye

Supportive validation

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Experimental details: Figure 2 Llgl1 regulates expression of cell lineage markers A. - D. , I. - L. Cells were incubated with the indicated cell lineage marker and sorted by FACS. A. - D. MCF12A shControl vs shLlgl1 (under normal growth conditions) were incubated with anti-CD49f-PE, anti-CD44-APC, and/or anti-CD24-FITC. E. and F. MCF12A shLlgl1 were sorted based on CD44/CD49f expression, CD444 hi /CD49f lo (E) and CD44 lo /CD49f hi (F). G. Protein lysates were isolated and analyzed by immunoblot using anti-Llgl1 and anti-betaactin antibodies. H. Protein lysates were analyzed by immunoblot using anti-Integrin alpha6 and anti-betaactin. (I-L) MCF10A shControl vs shLlgl1 (under normal growth conditions) were incubated with anti-CD49f-PE, anti-CD44-APC, and/or anti-CD24-FITC.
Conjugate: Green dye

Supportive validation

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Experimental details: Fig. 7 HOXC8 knockdown increase CSC proliferation and transformed phenotype. a Knockdown of HOXC8 was induced by lentiviral transduction of MCF10A cells with pLKO-Tet-HOXC8 vector and gene expression measured by TaqMan(r) qRT-PCR. Results are presented as relative fold expression relative to RPLP0 and pLKO-Tet-Scrambled vector used as calibrator ( n = 3). Results were analysed by Unpaired Student's t -test ( n = 3). *** P < 0.001. Bottom panel shows western blotting of nuclear lysates from cells transduced with pLKO-Tet-HOXC8 vector or pLKO-Tet-Scrambled vector was conducted to detect the expression of HOXC8 (34 kDa) and LAMIN A/C (41-50 kDa) as loading control. b Percentage of CD44 + /CD24 low/- , CD44 + /CD24 + , CD44 - /CD24 + cells after HOXC8 knockdown as measured by FACS analysis. Results were analysed by comparing cell populations levels in HOXC8 shRNA transduced cells compared to Scrambled shRNA ( n = 3). Statistical analysis was performed by Unpaired Student's t -test. * P < 0.05, ** P < 0.01. Representative FACS profiles of cells double stained with CD44-FITC and CD244-PE to analyse the CSC and non-CSC populations are also shown. c Mammospheres morphology and number obtained by self-renewal of cell after HOXC8 knockdown compared to scrambled control vector after 3 generations. Mammospheres numbers were compared by Unpaired Student's t -test ( n = 3). ** P < 0.01. Only spheres >= 100 muM were counted, images were taken at 10X magnification. d Colonies
Conjugate: Green dye

Supportive validation

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Experimental details: Fig. 8 HOXC8 downregulation prevents differentiation of MCF10A cells induced by retinoic acid. a Representative FACS profiles and percentage of CD44 + /CD24 low/- , CD44 + /CD24 + , CD44 - /CD24 + cells after retinoic acid (ATRA) treatment for 4 days and HOXC8 knockdown induced by doxycyclin ( n = 3). Statistical analysis was performed by One-way ANOVA followed by Bonferroni's multiple comparisons test. * P < 0.05, ** P < 0.01, *** P < 0.001. b Representative FACS profiles and percentage of CD44 + /CD24 low/- , CD44 + /CD24 + , CD44 - /CD24 + cells after retinoic acid (ATRA) treatment for 7 days and HOXC8 knockdown induced by doxycyclin ( n = 3). Statistical analysis was performed by One-way ANOVA followed by Bonferroni's multiple comparisons test. * P < 0.05, ** P < 0.01, *** P < 0.001
Conjugate: Green dye

Supportive validation

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Experimental details: Figure 1 Characterization of BCSC5 in vitro and in vivo. ( A ) Representative pictures of BCSC5 cultured in 3D and 2D conditions, scale bar 100 mum. ( B ) Sphere-forming capacity of BCSC5 cells in an anchorage-independent growth assay ( n = 3). Data represent means + SEM. ( C , D ) Expression patterns of CD24 and CD44 ( C ) as well as EpCAM and CD49f ( D ) in BCSC5 cells analyzed by flow cytometry. ( E ) Tumor formation in limiting dilution xenografts of BCSC5. ( F ) Representative growth curves for limiting dilution assay of BCSC5 xenografts in immunocompromised NOD/SCID mice. ( G ) Immunohistochemical (IHC) analysis of ER, PR, and HER2 on sections of the BCSC5 xenograft tumors, scale bar 100 mum.
Conjugate: Green dye

Supportive validation

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Experimental details: Figure 5 CD44 + /CD24 - levels detection in MCF10A and MDA-MB-231 cells with flow cytometry assay. ( A ). CD44 + /CD24 - evaluation in HMGB3-treated MCF10A cells. ( B ). CD44 + /CD24 - evaluation in siHMGB3-treated MDA-MB-231 cells. * p
Conjugate: Green dye

Supportive validation

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Experimental details: Figure 10 Determination for CD44 + /CD24 - levels in HMGB3/siHIF1alpha-treated MCF10A cells and siHMGB3/HIF1alpha-treated MDA-MB-231 cells using flow cytometry assay. ( A ). CD44 + /CD24 - evaluation in HMGB3/siHIF1alpha-treated MCF10A cells. ( B ). CD44 + /CD24 - evaluation in siHMGB3/HIF1alpha-treated MDA-MB-231 cells. * p
Conjugate: Green dye