anti cd24 mab rabbit  (Proteintech)

Journal: eLife

Article Title: Proliferative exhausted CD8+ T cells exacerbate long-lasting anti-tumor effects in human papillomavirus-positive head and neck squamous cell carcinoma

doi: 10.7554/elife.82705

Figure Lengend Snippet: Figure 5. The expression of CDK4 gene in P-Tex2 cluster is associated with the treatment outcomes of HPV+ head and neck squamous cell carcinoma (HNSCC) patients. (a) Single-cell transcriptomic profiling of HNSCC tumor microenvironment (TME). Twenty cell clusters are identified, colored by cell types. (b) The proliferation status of P-Tex and epithelial cells in violin plot. (c) The kernel density estimate distribution of proliferation markers (CDK4 and MKI67) and epithelial cancer cell markers (KRT15 and CD24) in uniform manifold approximation and projection (UMAP) plots. (d) The overall survival rate of HPV+/HPV- HNSCC patients in The Cancer Genome Atlas (TCGA) cohort related to the expression levels of CDK4 gene, adjusted for age and gender. (e) The proportion of P-Texs, T-Tex, and TEFF clusters in HPV+ and HPV- samples in TCGA cohort by using the deconvolution algorithm; statistics were assessed by Chi-square tests. Marker genes that were used to define cell clusters in (a) are deconvolved into the TCGA data to obtain the proportion of P-Texs, T-Tex, and Teff clusters in the TCGA cohort. (f) The cell viability of P-Tex and cancer epithelial cells assessed by CCK8 experiment after Abemaciclib treated in vitro. ***: p<0.001, **: p<0.01, *: p<0.05.

Article Snippet: Continued on next page Key resources table Reagent type (species) or resource Designation Source or reference Identifiers Additional information Sequence- based reagent Chromium Single Cell 5’ Gel Bead and Library Construction Kit 10× Genomics PN- 1000006; PN- 1000020 Sequence- based reagent Single Cell V(D)J Enrichment Kit Human T cell 10× Genomics PN- 1000005 Sequence- based reagent Visum Spatial Library Construction kit 10× Genomics PN- 1000184 Antibody FITC Mouse Anti- Human CD3 (mouse monoclonal) BD Pharmingen Cat#555332 FACS (0.8 μL per test) Antibody PE anti- human CD8a Antibody (mouse monoclonal) Cell Signaling Technology Cat#300908, Clone: HIT8a FACS (5 μL per test) Antibody Anti- human CD279 (PD- 1) (mouse monoclonal) Biolegend Cat#329920 FACS (5 μL per test) Antibody UBE2C (B- 8) PE (mouse monoclonal) Santacruz Cat#Sc271050 FACS (1:100) Antibody PE/Cyanine7 anti- human CD161 (mouse monoclonal) Biolegend Cat#339917 FACS (5 μL per test) Antibody Anti- CD8α (mouse monoclonal) Cell Signaling Technology Cat#70306S IHC (1:200) Antibody Anti- PD- 1 (Rabbit monoclonal) Cell Signaling Technology Cat#84,651T IHC (1:200) Antibody Anti- Ki67 (Rabbit monoclonal) Abcam Cat#ab16667, Clone: SP6 IHC (1:50) Antibody Anti- CDK4 (Rabbit monoclonal) Cell Signaling Technology Cat#12790 IHC (1:1000) Antibody Anti- CD24 (Rabbit Polyclonal) Proteintech Cat#10600- 1- AP IHC (1:200) Antibody Anti- HLA- DR (Rabbit monoclonal) Abcam Cat#ab92511 IHC (1:200) Antibody Anti- EpCAM (Rabbit monoclonal) Abcam Cat#ab223582, Clone: EPR20532- 225 IHC (1:500) Commercial assay or kit Cell Counting Kit- 8 MedChemExpress Cat#HY- K0301 Commercial assay or kit Manual Opal 7- Color IHC Kit Akoyabio Cat#NEL811001KT Cell line (Homo sapiens) Laryngeal carcinoma This paper FD- LSC- 1 Cell line maintained in State Key Laboratory of Biotherapy, West China Medical School, Sichuan University Chemical compound, drug Abemaciclib (LY2835219) Topscience Cat#T2381 Software, algorithm Cell Ranger count 10× Genomics v3.1.0 Software, algorithm Cell Ranger VDJ 10× Genomics v6.1.1 Software, algorithm Space Ranger 10× Genomics v2.0 Software, algorithm Seurat R RRID:SCR_007322 Software, algorithm monocle3 R RRID:SCR_018685 Software, algorithm CellChat R RRID:SCR_021946 Cheng, Qiu, Rao et al. eLife 2023;12:e82705.

Techniques: Expressing, Marker, In Vitro

Journal: PLoS ONE

Article Title: Cell Atavistic Transition: Paired Box 2 Re-Expression Occurs in Mature Tubular Epithelial Cells during Acute Kidney Injury and Is Regulated by Angiotensin II

doi: 10.1371/journal.pone.0093563

Figure Lengend Snippet: The RT-PCR (A) and western blot (B) data showed that 10 −9 mol/L of Ang II could stimulate CD24 expression in NRK-52E cells shortly after the expression of Pax2; Ang II induced CD24 expression was started from 1 h post exposure, reached a peak at 4 h and began to disappear at 6 h (*P<0.05, compared with Time = 0 h). (C) Immunofluorescence analyses showed that CD24 was expressed at the surface of NRK-52E cells.

Article Snippet: The following antibodies were used; rabbit anti-rat Pax2 polyclonal antibody (1∶400, Santa Cruz), rabbit anti-rat CD24 polyclonal antibody (1∶200, Santa Cruz), anti-mouse IgG and anti-rabbit IgG secondary antibody (1∶1000, Cell signaling technology).

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Immunofluorescence

Journal: PLoS ONE

Article Title: Cell Atavistic Transition: Paired Box 2 Re-Expression Occurs in Mature Tubular Epithelial Cells during Acute Kidney Injury and Is Regulated by Angiotensin II

doi: 10.1371/journal.pone.0093563

Figure Lengend Snippet: NRK-52E cells were pretreated with PD123319 (10 −6 mol/L) and the AG490 (10 −6 mol/L) for 10 min before Ang II (10 −9 mol/L) administration. Cells were harvest at 4 h post exposure for RT-PCR and western blot assays. Results of RT-PCR (A) and western blot (B) showed that both the AT2R blocker PD123319 and JAK2 blocker AG490 could block the stimulatory effect of Ang II on CD24 expression in NRK-52E cells (*P<0.05).

Article Snippet: The following antibodies were used; rabbit anti-rat Pax2 polyclonal antibody (1∶400, Santa Cruz), rabbit anti-rat CD24 polyclonal antibody (1∶200, Santa Cruz), anti-mouse IgG and anti-rabbit IgG secondary antibody (1∶1000, Cell signaling technology).

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Blocking Assay, Expressing

Journal: PLoS ONE

Article Title: Cell Atavistic Transition: Paired Box 2 Re-Expression Occurs in Mature Tubular Epithelial Cells during Acute Kidney Injury and Is Regulated by Angiotensin II

doi: 10.1371/journal.pone.0093563

Figure Lengend Snippet: NRK-52E cells were transfected with Pax2 shRNA or emptily plasmid before Ang II administration. Cells were harvest at 4 h post exposure. RT-PCR (A) and western blot (B) results showed that knockdown of Pax2 significantly suppressed Ang II induced CD24 expression in NRK-52E cells, while transfection with emptily plasmid pGenesil-1 did not affect CD24 expression.

Article Snippet: The following antibodies were used; rabbit anti-rat Pax2 polyclonal antibody (1∶400, Santa Cruz), rabbit anti-rat CD24 polyclonal antibody (1∶200, Santa Cruz), anti-mouse IgG and anti-rabbit IgG secondary antibody (1∶1000, Cell signaling technology).

Techniques: Transfection, shRNA, Plasmid Preparation, Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing

Journal: Stem Cells International

Article Title: Expression of CD24 in Human Bone Marrow-Derived Mesenchymal Stromal Cells Is Regulated by TGF β 3 and Induces a Myofibroblast-Like Genotype

doi: 10.1155/2016/1319578

Figure Lengend Snippet: Analysis of CD24 expression in hBMSCs. (a) hBMSC mRNA from 18 different donors in passage 2 of in vitro culture were analyzed for the expression of CD24 mRNA by qRT-PCR and analyzed for gender-dependent differences in mRNA expression levels. The relative expression values are plotted as fold change in mRNA expression (ΔΔCt) relative to the sample with the lowest relative CD24 mRNA expression. (b) The mRNA expression of CD24 was analyzed from passage 1 to passage 4 of in vitro culture in 4 individual donors. The expression values are normalized to the expression levels of the respective donor in passage 1 (ΔΔCt). (c) The expression of CD24 protein was evaluated by western blot with a polyclonal rabbit anti-CD24 antibody. In order to determine which bands were specific for CD24 hBMSCs were transduced with either a shRNA directed against CD24 mRNA (CD24 Down) or a scrambled shRNA (Control). Cell extracts from these two groups were then compared for CD24 reactive bands: the western blot bands with reduced intensity in the CD24 Down Group are marked with black arrows and represent bands specific for CD24. β -actin was used as loading control.

Article Snippet: Appropriate primary antibodies (monoclonal mouse anti-CD24 antibody (ML5) (BioLegend), polyclonal rabbit anti-CD24 antibody (FL-80, Santa Cruz Biotechnology, Dallas, USA), and monoclonal mouse anti-alpha smooth muscle actin (R&D Systems, Minneapolis, Minnesota, USA)) were used at a concentration of 1 μ g/mL in 1% (w/v) BSA and 0.1% (w/v) Triton X-100 in PBS and incubated for 1 hour at room temperature.

Techniques: Expressing, In Vitro, Quantitative RT-PCR, Western Blot, Transduction, shRNA

Journal: Stem Cells International

Article Title: Expression of CD24 in Human Bone Marrow-Derived Mesenchymal Stromal Cells Is Regulated by TGF β 3 and Induces a Myofibroblast-Like Genotype

doi: 10.1155/2016/1319578

Figure Lengend Snippet: Cellular localisation of CD24. (a), (b), and (c) hBMSCs were analyzed by flow cytometry for the expression of CD24 protein. An exemplary histogram depicting the low extracellular CD24 expression can be found in (a); an exemplary histogram depicting the high intracellular CD24 protein density is shown in (b). (c) shows the percent expression of CD24 protein extra- and intracellularly in three independent experiments with hBMSCs from different donors. (d) Immunocytochemical analysis of the intracellular CD24 protein expression showing a diffuse cytosolic and a strong nuclear reactivity. A z-stack image is shown to highlight the nuclear localisation of CD24 (monoclonal anti-CD24 antibody, clone ML-5). Green = CD24. Blue = nuclear staining. Scale: 20 μ m. A high resolution image of (d) can be found in Supplemental Figure 5.

Article Snippet: Appropriate primary antibodies (monoclonal mouse anti-CD24 antibody (ML5) (BioLegend), polyclonal rabbit anti-CD24 antibody (FL-80, Santa Cruz Biotechnology, Dallas, USA), and monoclonal mouse anti-alpha smooth muscle actin (R&D Systems, Minneapolis, Minnesota, USA)) were used at a concentration of 1 μ g/mL in 1% (w/v) BSA and 0.1% (w/v) Triton X-100 in PBS and incubated for 1 hour at room temperature.

Techniques: Flow Cytometry, Expressing, Staining

Journal: Stem Cells International

Article Title: Expression of CD24 in Human Bone Marrow-Derived Mesenchymal Stromal Cells Is Regulated by TGF β 3 and Induces a Myofibroblast-Like Genotype

doi: 10.1155/2016/1319578

Figure Lengend Snippet: Reciprocal regulation of CD24 and TGF β 3. (a) qRT-PCR analysis of CD24 mRNA expression after stimulation with 10 ng/mL TGF β 3 for 4 h, 24 h, and 96 h relative to unstimulated control cells (ΔΔCt). (b) Exemplary histogram showing induction of CD24 surface expression after treatment with 10 ng/mL TGF β 3 or with 2.5 ng/mL TGF β 1 for 7 days as analyzed by flow cytometry. (c) Flow cytometric analysis of CD24 surface protein expression on hBMSCs after 7 days in unstimulated control cells (Control), in cells treated with 10 ng/mL TGF β 3 (TGF β 3), in cells treated with 10 ng/mL TGF β 3 + 20 μ M SB431542 (SB431542), or in cells treated with 10 ng/mL TGF β 3 and 4 μ g/mL anti-TGF β 3 antibody (anti-TGF β 3). (d) and (e) qRT-PCR analysis of TGF β 3 (d) and CD24 (e) mRNA expression after knockdown (CD24 Down) or overexpression (CD24 Up) of CD24 in hBMSCs after 7 days relative to respective control hBMSCs ( n = 3 independent experiments with hBMSCs from different donors).

Article Snippet: Appropriate primary antibodies (monoclonal mouse anti-CD24 antibody (ML5) (BioLegend), polyclonal rabbit anti-CD24 antibody (FL-80, Santa Cruz Biotechnology, Dallas, USA), and monoclonal mouse anti-alpha smooth muscle actin (R&D Systems, Minneapolis, Minnesota, USA)) were used at a concentration of 1 μ g/mL in 1% (w/v) BSA and 0.1% (w/v) Triton X-100 in PBS and incubated for 1 hour at room temperature.

Techniques: Quantitative RT-PCR, Expressing, Flow Cytometry, Over Expression

Journal: Stem Cells International

Article Title: Expression of CD24 in Human Bone Marrow-Derived Mesenchymal Stromal Cells Is Regulated by TGF β 3 and Induces a Myofibroblast-Like Genotype

doi: 10.1155/2016/1319578

Figure Lengend Snippet: IPA upstream regulator analysis. Molecules resulting in similar changes in the pattern of gene expression as the overexpression of CD24 or the knockdown of CD24 are designated with high activational z -score values. Listed are those molecules with a z -score higher than 2 while exhibiting opposing algebraic signs in the CD24 Up and CD24 Down Groups.

Article Snippet: Appropriate primary antibodies (monoclonal mouse anti-CD24 antibody (ML5) (BioLegend), polyclonal rabbit anti-CD24 antibody (FL-80, Santa Cruz Biotechnology, Dallas, USA), and monoclonal mouse anti-alpha smooth muscle actin (R&D Systems, Minneapolis, Minnesota, USA)) were used at a concentration of 1 μ g/mL in 1% (w/v) BSA and 0.1% (w/v) Triton X-100 in PBS and incubated for 1 hour at room temperature.

Techniques: Expressing, Over Expression, Infection, Activation Assay

Journal: Stem Cells International

Article Title: Expression of CD24 in Human Bone Marrow-Derived Mesenchymal Stromal Cells Is Regulated by TGF β 3 and Induces a Myofibroblast-Like Genotype

doi: 10.1155/2016/1319578

Figure Lengend Snippet: Biological functions influenced by CD24 and TGF β 3. The top five canonical pathways regulated by knockdown of CD24 (CD24 Down), overexpression of CD24 (CD24 Up), or stimulation with TGF β 3 (TGF β 3) in hBMSCs based on their significance ( P value) calculated by Ingenuity Pathway Analysis using the right-tailed Fisher's Exact Test using the entire data set. The dotted line is the threshold value with P = 0.05.

Article Snippet: Appropriate primary antibodies (monoclonal mouse anti-CD24 antibody (ML5) (BioLegend), polyclonal rabbit anti-CD24 antibody (FL-80, Santa Cruz Biotechnology, Dallas, USA), and monoclonal mouse anti-alpha smooth muscle actin (R&D Systems, Minneapolis, Minnesota, USA)) were used at a concentration of 1 μ g/mL in 1% (w/v) BSA and 0.1% (w/v) Triton X-100 in PBS and incubated for 1 hour at room temperature.

Techniques: Over Expression

Journal: Stem Cells International

Article Title: Expression of CD24 in Human Bone Marrow-Derived Mesenchymal Stromal Cells Is Regulated by TGF β 3 and Induces a Myofibroblast-Like Genotype

doi: 10.1155/2016/1319578

Figure Lengend Snippet: Expression of alpha smooth muscle actin. Intracellular immunocytochemical staining of alpha smooth muscle actin after overexpression of CD24 (CD24, left column) or truncated CD34 as control (CD34, right column). Alpha smooth muscle actin: staining of alpha smooth muscle actin (green), and nuclear staining (blue) show that in hBMSCs overexpressing CD24 reactivity for alpha smooth muscle actin increases relative to control cells (Scale 50 μ m). CD24: staining of CD24 (green), and nuclear staining (blue) verify that transduction rate of CD24 is nearly complete (Scale 100 μ m). CD34: staining of CD34 (green), and nuclear staining (blue) verify that control cells were transduced efficiently with truncated CD34 (Scale 100 μ m). Secondary antibody control: staining control with secondary antibody only (green) and nuclear staining (blue) (Scale 50 μ m).

Article Snippet: Appropriate primary antibodies (monoclonal mouse anti-CD24 antibody (ML5) (BioLegend), polyclonal rabbit anti-CD24 antibody (FL-80, Santa Cruz Biotechnology, Dallas, USA), and monoclonal mouse anti-alpha smooth muscle actin (R&D Systems, Minneapolis, Minnesota, USA)) were used at a concentration of 1 μ g/mL in 1% (w/v) BSA and 0.1% (w/v) Triton X-100 in PBS and incubated for 1 hour at room temperature.

Techniques: Expressing, Staining, Over Expression, Transduction