12-9459-42

antibody from Invitrogen Antibodies

Targeting: PTPRC CD45, GP180, LCA, T200

Provider product page for 12-9459-42

Flow cytometry

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Antibody data

Antibody Data
Antigen structure
References [31]
Comments [0]
Validations
Flow cytometry [3]
Other assay [13]

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Product number: 12-9459-42 - Provider product page
Provider: Invitrogen Antibodies
Product name: CD45 Monoclonal Antibody (2D1), PE, eBioscience™
Antibody type: Monoclonal
Antigen: Other
Description: Description: The 2D1 monoclonal antibody reacts with all isoforms of human CD45, also known as Leukocyte Common Antigen (LCA). CD45 is expressed by all hematopoietic cells excluding circulating erythrocytes and platelets. The cytoplasmic portion of CD45 has tyrosine phosphatase enzymatic activity and plays an important role in activation of lymphocytes. Applications Reported: This 2D1 antibody has been reported for use in flow cytometric analysis. Applications Tested: This 2D1 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: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered.
Reactivity: Human
Host: Mouse
Conjugate: Yellow dye
Isotype: IgG
Antibody clone number: 2D1
Vial size: 100 Tests
Concentration: 5 µL/Test
Storage: 4° C, store in dark, DO NOT FREEZE!

PTPRC protein structure - 12-9459-42 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Staining of normal human peripheral blood cells with Mouse IgG1 K Isotype Control PE (Product # 12-4714-81) (blue histogram) or Anti-Human CD45 PE (purple histogram). Cells in the lymphocyte gate were used for analysis.
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of CD45 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR664203_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Flow cytometry analysis of CD45 was performed by staining THP-1 CD45 Knock out cells with 0.25 µg Mouse IgG1 kappa Isotype Control (P3.6.2.8.1), PE, eBioscience™ (Product # 12-4714-82) or 0.25 µg CD45 Monoclonal Antibody (2D1), PE, eBioscience™ (Product # 12-9459-42, blue histogram). THP-1 Cas9 control cells were also stained with0.25 µg CD45 Monoclonal Antibody (2D1), PE, eBioscience™ (Product # 12-9459-42, pink histogram). Lossof signal was observed in the KOcells stained with CD45 antibody clone 2D1 but not in the control Cas9cells. Viable cells were used for analysis, as determined by Fixable Viability DyeeFluor™780 (Product # 65-0865-18).
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Staining of normal human peripheral blood cells with Mouse IgG1 K Isotype Control PE (Product # 12-4714-81) (blue histogram) or Anti-Human CD45 PE (purple histogram). Cells in the lymphocyte gate were used for analysis.
Conjugate: Yellow dye

Supportive validation

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

Supportive validation

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

Supportive validation

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

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 2 In silico validation of the immune cell scoring method. a In silico validation of immune cell scores using simulated mixing proportions. RNA-Seq profiles of FACS-sorted NK cells, macrophages, CD 4 + and CD 8 + T cells, and non-immune CD 45 - cells were mixed with known proportions to obtain a ""clean"" mixture. Noise was added at varying SNRs. Mixing levels were then inferred by ssGSEA from the ""clean"" and noisy mixtures. The Spearman correlations between the simulated and inferred levels ( top panel ) and the bootstrap p values for these correlation values ( bottom panel ) are shown on the y-axes (Additional file 1 : Figure S18 and "" Methods "" for the calculation of the bootstrap p values). b Validation of IIS with methylation-based leukocyte fractions. Spearman correlations between the two orthogonal scores are shown on the x-axis for 13 tumor types. c Validation of TIS with TCR beta chain abundance. Both scores are computationally inferred from RNA-Seq data but employ different approaches to measure T cell levels. Spearman correlations are shown on the x-axis for 19 tumor types
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2. EV quantification and cell origin in association with HIV. Purified plasma EVs fromuninfected (Control, n = 8) and HIV+ patients (n = 17) were stained with the lipophilic fluorescent tracer dye DiD to count total vesicles ( Figure 2A and B ), then labeled with antibodies directed against receptors CD45 ( Figure 2C and D ), CD4 ( Figure 2E and F ), and CD8 ( Figure 2 G and H ) to evaluate cell origin by cytofluorometry. Asterisks denote significant difference (** P
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 1 Characterization of young and aged MSCs and exosomes. (A) Surface marker profiling of young-MSCs and aged-MSCs. (B) SA-beta-Gal staining showed that senescence increased significantly in aged MSCs. (C) Representative immunoblot images and quantitative analysis of p21, p53, and p16 protein level in young and aged-MSCs. ( n = 3). (D) Quantitation of cell cycle phases by propidium iodide staining. ( n = 3). (E) The CCK-8 assay showed that aged MSCs grew more slowly than young MSCs. ( n = 6). (F) Young and aged exosomes were observed using TEM. (G) The exosome surface markers were analyzed by Western blot. (H) Nanoparticle tracking analysis was used to analyze the particle size and concentration of Young-Exo and Aged-Exo. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; NS, not significant.
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 1 Culture and identification of bone marrow mesenchymal stem cells. (A) Light microscopy of BMSCs (scale bar=100 um for the left image and 200 um for the right image). (B) Percentage of CD34-, CD45-, CD73-, CD90- and CD105-positive BMSCs were detected by flow cytometry. BMSC, bone marrow mesenchymal stem cell.
Conjugate: Yellow dye

Supportive validation

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Experimental details: Figure 1. Morphology and immune phenotype of adipose-derived stem cells (ADSCs) were identified by morphological observation and flow cytometry. (a) Morphology of the primary (P1) and third passage (P3) of ADSCs. Images were acquired at 200x magnification. (b) Immune phenotype of ADSCs. The average data from three independent experiments were shown as mean +- standard deviation
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 4 oMG expressed microglia-characteristic cell surface markers and showed similar functional immune and phagocytic properties as adult MG. a Flow cytometric analyses of the expression pattern of microglial extracellular markers on CD11b+-gated oMG (oMG 1, 3, and 5) compared to adult MG derived from three separate brain regions from adult MG1.1. (eight organoids were pooled per donor (oMG 1, 3, and 5) after 52 days in culture). b Morphology of magnetic automated cell sorted CD11b+ oMG 1 and adult MG in bright field microscope after 1 week in culture. Scale bar 40 mum. c mRNA expression, determined by qRT-PCR, of pro-inflammatory cytokines IL6 and IL1B after 6 h stimulation with LPS was significantly higher in oMG compared to adult MG (Mann-Whitney test IL6 and IL1B: U = 0, n = 4, p = 0.03). LPS-stimulated response relative to control condition without LPS. ( n = 4 experiments, eight organoids pooled per experiment; adult MG1.1) (* p < 0.05). d Anti-inflammatory response of oMG and adult MG was compared by qRT-PCR for expression of anti-inflammatory genes CD163 and MRC1 upon 72 h stimulation with dexamethasone. Dexamethasone-stimulated response relative to control condition without dexamethasone. (oMG, n = 3 separate experiments in which oMG were isolated from > 4 pooled cerebral organoids from iPSC 1 per experiment; adult MG, n = 4). e Phagocytosis capacity was tested oMG 1 and adult MG by performing a phagocytosis assay with iC3b-coated green-yellow
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: 01 Figure S1 Isotype control for TREM-1 and TREM-2 expression in monocytes and neutrophils of the study subjects. Cell phenotype was performed using anti-human antibodies and isotypes for monocytes (CD45+, CD14+, HLA-DR+) represented in figure A and neutrophils (CD45+, CD16+) in figure B. A) The representative figures for isotype control for monocytes gating; live monocytes were gated from FSC/SSC gating. Then CD45+ were gated using the basal level of CD45 Isotype and analyzed for CD14 and HLA-DR expression with respective CD14 and HLA-DR Isotypes. The double positive populations (CD14+ HLA-DR+) were further gated for expression of TREM-1+ and TREM-2+ using the basal level of TREM-1 and TREM-2 Isotypes. B) The representative figures for Isotype control for neutrophils gating; live granulocytes were gated from FSC/SSC gating. Then CD45+ were gated using the basal level of CD45 Isotype and analyzed for CD16 expression using the basal level of CD16 Isotype. CD16+ populations were further gated for expression of TREM-1 and TREM-2 using the basal level of TREM-1 and TREM-2 Isotypes. Figure S2 Hematoxylin and Eosin staining for fatty liver grading and inflammation. H & E in biopsy samples of SO - D - , SO + D - and SO + D + groups respectively in liver (images Aa, Ab & Ac), omental fat (images Ba, Bb & Bc) and subcutaneous fat (images Ca, Cb & Cc). Liver biopsy samples showed steatosis in SO + D - and fibrosis and cirrhosis in SO + D + groups. Size of the adipocyte was larger in su
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 1. Characterization of PDLSCs. (A) PDL cell clusters exhibited radiating or whirlpool-like morphology. The central structure in this image is a fragment of PDL tissue. Scale bar, 200 mum (B) CD146 + PDLSCs were small, round, fusiform and triangular. Scale bar, 100 mum. (C) PDLSCs were positive for the stem cell markers CD44, CD90 and CD105, but negative for CD34 and CD45, as detected by flow cytometry. PDL, periodontal ligament; PDLSCs, PDL stem cells.
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig 3 Cancer cell lines and mf significantly upregulates the cell surface expressions of PDL1, PDL2, CD206 and VCAM-1 on human monocytes. Human monocytes were cultured in media alone, or with CMFDA-labeled three different cancer cell lines (MDA, OVCAR, U87), or with live mf of Brugia malayi for 48hr. Cells were harvested and cell surface expression PDL1, PDL2, CD206, VCAM-1, and CD163 was measured using flow cytometry gated on CD45 + /CMFDA - monocytes. (A) One representative set (n = 15) of flow histograms demonstrating cell surface expression in unexposed human monocytes and after exposure to mf or different cancer cell lines. (B). The data are expressed as the geometric mean with 95% confidence interval of the mean fluorescent intensity of unexposed and exposed monocytes ( n = 15). * P< 0.05, ** P< 0.005.
Conjugate: Yellow dye

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 4 Comparison of MSCs derived from 2D-hESCs and 3D-hESCs. a The morphology of 2D- and 3D-hESC-MSCs at different stages of differentiation. b Flow cytometry analysis revealed specific MSC surface markers (CD44, CD29, and CD105) with negative controls (CD34, CD19, and CD45) in 2D- and 3D-hESC-MSCs. c Immunostaining of differentiated 3D-hESC-MSCs expressing an adipocyte marker (FABP-4), osteocytes maker (osteocalcin), and chondrocytes marker (aggrecan)
Conjugate: Yellow dye