MA5-11828

antibody from Invitrogen Antibodies

Targeting: CD24 CD24A

Provider product page for MA5-11828

Western blot

Antibody data

Antibody Data
Antigen structure
References [38]
Comments [0]
Validations
Western blot [1]
Immunocytochemistry [3]
Immunohistochemistry [1]
Flow cytometry [3]
Other assay [6]

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Product number: MA5-11828 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD24 Monoclonal Antibody (SN3)
Antibody type: Monoclonal
Antigen: Other
Description: MA5-11828 targets CD24 in FACS, IHC (F), ICC/IF and WB applications and shows reactivity with Human and Mouse samples. This antibody detects a few non-specific bands in BAF-3 cell lysates.
Antibody clone number: SN3
Concentration: 0.2 mg/mL

CD24 protein structure - MA5-11828 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of CD24 (green) showing staining in the cytoplasm of MCF-7 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CD24 monoclonal antibody (Product # MA5-11828) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of CD24 (green) showing staining in the cytoplasm of NIH-3T3 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CD24 monoclonal antibody (Product # MA5-11828) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of CD24 (green) showing staining in the cytoplasm of U251 cells (right) compared to a negative control without primary antibody (left). Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with a CD24 monoclonal antibody (Product # MA5-11828) in 3% BSA-PBS at a dilution of 1:20 and incubated overnight at 4 ºC in a humidified chamber. Cells were washed with PBST and incubated with a DyLight-conjugated secondary antibody in PBS at room temperature in the dark. F-actin (red) was stained with a fluorescent red phalloidin and nuclei (blue) were stained with Hoechst or DAPI. Images were taken at a magnification of 60x.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of CD24 in BAF-3 cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde, washed with PBS, and incubated with CD24 monoclonal antibody (Product # MA5-11828) at a dilution of 0.25 µg/test for 60 min at room temperature. Cells were then blocked in a solution of 2% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody, and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of CD24 in PBMC cells compared to an isotype control (blue). Human blood was collected, combined with a hydrophilic polysaccharide, centrifuged, transferred to a conical tube and washed with PBS. 50 µL of cell solution was added to each tube at a dilution of 2x10^7 cells/mL, followed by the addition of 50 µL of isotype control and primary antibody (Product # MA5-11828) at a dilution of 0.5 µg/test. Cells were incubated for 30 min at 4°C and washed with a cell buffer, followed by incubation with a DyLight 488-conjugated goat anti-mouse IgG (H+L) secondary for 30 min at 4°C in the dark. FACS analysis was performed using 400 µL of cell buffer.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of CD24 in PBMC cells compared to an isotype control (blue). Human blood was collected, combined with a hydrophilic polysaccharide, centrifuged, transferred to a conical tube and washed with PBS. 50 µL of cell solution was added to each tube at a dilution of 2x10^7 cells/mL, followed by the addition of 50 µL of isotype control and primary antibody (Product # MA5-11828) at a dilution of 0.5 µg/test. Cells were incubated for 30 min at 4°C and washed with a cell buffer, followed by incubation with a DyLight 488-conjugated goat anti-mouse IgG (H+L) secondary for 30 min at 4°C in the dark. FACS analysis was performed using 400 µL of cell buffer.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: NULL

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 CD44v3-10 expression is positively correlated with poor prognosis in breast tumors. ( a ) Representative immunofluorescence analyses of total CD44, CD44v and CD24 in human breast cancer samples. White and yellow arrows denote CD24 - /CD44 + /CD44v + and CD24 - /CD44 + /CD44v - cells, respectively. Insets in white and yellow boxes show representative CD24 - /CD44 + /CD44v + and CD24 - /CD44 + /CD44v - cells, respectively. ( b ) CD44v3-10 and CD44s expression ratios analyzed by qPCR in human breast cancer samples, results are expressed as mean+-S.D., n =3. ( c ) Distant metastasis-free survival (DMFS) analysis of the patients in the KM-Plotter database stratified by ESRP1 expression, ( n =1610). ( d ) Correlation of ESRP1 expression and CD44v3-10/CD44s expression ratios in Qilu clinical samples ( n =45). ( e-g ) Distant metastasis-free survival (DMFS) analysis of Qilu clinical samples stratified by expression levels of CD44v3-10 ( e ), total CD44 ( f ) and CD44s ( g ) ( n =63). Scale bars, 10 mu m (insets) and 20 mu m (others)

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 3 Analysis of CD24 expression in human neuroblastoma cells. A) Schematic of the alignment of CD24 splice variants 1 and 7. B & C) Absolute quantification of CD24 expression by quantitative real-time PCR of total RNA acquired from neuroblastoma cells, measuring CD24 splice variants 1 (B) and 7 (C). Copy number values were normalized to the corresponding GAPDH values to determine the relative copy number. ** p > 0.05, Student's t-test. D) Western blot analysis of CD24 expression in the total cell lysates of neuroblastoma cells. GAPDH was used as a loading control. All results are representative of the combined data of experiments performed in triplicate, with error bars representing standard deviation.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 4 Analysis of the role of CD24 in Zika-virus infected neuroblastoma cells. A) Western blot analysis of siRNA-mediated knock-down of CD24 expression in IMR-32 cells. Samples include Negative Control siRNA and CD24 siRNA. B) Western blot analysis of the expression of Envelope protein and NS1 (Non-Structural 1) protein in IMR-32 cells after siRNA-mediated knock-down of CD24 expression, 96 hours after Zika infection (MOI = 10). Samples include control cells treated with non-infected conditioned media and infected IMR-32 cells transfected with either Negative Control siRNA or CD24 siRNA. C) Western blot analysis of CD24 expression in the human neuroblastoma cell line SK-N-AS, comparing wild type (WT) to stably selected ""Vector Only"" (VO), CD24 variant 1 (V1), and CD24 variant 7 (V7). D) Western blot analysis of Zika NS1 protein expression 96 hours after Zika infection in CD24-stably expressing SK-N-AS cells, comparing wild type (WT) to stably selected Vector Only (VO), CD24 variant 1 (V1), and CD24 variant 7 (V7). GAPDH was used as a load control for all experiments. All results are representative of the combined data of experiments performed in triplicate.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 3 TGEV infection results in CD24 + SSC high cells (Paneth cells) loss and Notch signaling inactivation. a Representative FACS of CD24 in TGEV-infected IPEC-J2 cells. b Quantification of CD24 + SSC high (Paneth cells) and CD24 + SSC low (enteroendocrine cells) number in TGEV-infected IPEC-J2 cells. c Jejunum from TGEV-infected piglets were analyzed by quantitative PCR (qPCR) for makers of ISCs niche signaling (Notch, Wnt, and EGF signaling). d Representative Notch factors mRNA change in TGEV-infected IPEC-J2 cells. e Western blot for Notch effector Hes5 and Notch ligand DII4 in jejunum from TGEV-infected piglets. f Quantitation of bands to demonstrate the protein level of DII4 and Hes5. g Representative Notch factors (DII4 and Hes5) and intestinal epithelial cells markers Muc2, SI, CgA, and CD24 were tested by western blot in TGEV-infected IPEC-J2 cells. h TGEV content was tested by FACS in 24 h post TGEV-infected IPEC-J2 cells, which were pre-treated by DAPT for 12 h before TGEV infection, and DAPT continued addition during TGEV infection period.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 6 TGEV-encoded NSP10 and NSP16 mediate cell proliferation and Lgr5 ISCs fate. a Cell proliferation (KI67), CD24 cells number and Notch signaling factors DII4 and Hes5 were analyzed in TGEV gene stable cell lines. Venn diagram depicts the numbers of decreased cell proliferation and CD24 cells number and down-regulated Notch signaling identified by using FACS and western blot. b FACS for KI67 in TGEV NSPs stable cell lines. c Notch factor and intestinal epithelium markers Muc2, SI, CgA, and CD24 were tested by western blot in TGEV NSPs stable cell lines. d FACS for CD24 in TGEV NSPs stable cell lines.