Ab40763

Manufactured by Abcam

Sourced in United States, United Kingdom

Ab40763 is a monoclonal antibody raised against a synthetic peptide derived from the N-terminus of human Amyloid Beta 40. The antibody is designed for research use only and its core function is to detect and bind to Amyloid Beta 40 in various sample types.

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Lab products found in correlation

Ab81289, by Abcam (4 mentions) Ab150077, by Abcam (3 mentions) Ab225, by Abcam (3 mentions) Ab16669, by Abcam (3 mentions) Ab189524, by Abcam (2 mentions) Ab133582, by Abcam (2 mentions) Ab202122, by Abcam (2 mentions) Ab9498, by Abcam (2 mentions) Inverted fluorescence microscope, by Leica (1 mentions) Cytoflex s flow cytometry, by Beckman Coulter (1 mentions) Adipogenic differentiation medium, by Cyagen (1 mentions) Ab189786, by Abcam (1 mentions) Ab76956, by Abcam (1 mentions) Ab229126, by Abcam (1 mentions) Ab92574, by Abcam (1 mentions) Polyvinylidene fluoride membrane, by Merck Group (1 mentions) Ab93876, by Abcam (1 mentions) Image pro plus 6, by Media Cybernetics (1 mentions) Sds lysis buffer, by Beyotime (1 mentions) Ecl808 25, by Biomiga (1 mentions)

Spelling variants of this product

Anti-human CD45 (ab40763; (1 citations)

20 protocols using ab40763

Isolation and Characterization of Mesenchymal Stem Cells

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MSCs were isolated according to the previous method [26 (link)]. In brief, bone marrow of the four limbs was obtained by flushing with 26-gauge syringe needle containing phosphate-buffered saline (PBS) under sterile conditions. The contents were washed and centrifuged at 1500 rpm for 5 min. And then MSCs was resuspended with mesenchymal stem cell medium (MSCM, Sciencell. USA) and maintained in a 37 ºC incubator with 5% CO2. MSCs were observed under a fluorescence inverted microscope (Leica. Germany) and passaged when they reached 80–90% confluence.
For phenotypic characterization, 5 × 10⁵ MSCs were trypsinized and washed twice with PBS, and were incubated with fluorescein isothiocyanate (FITC)-conjugated anti-CD29 (ab255354), anti-CD44 (ab189524) and anti-CD45 (ab40763) (all from Abcam, USA) at a dilution rate of 1: 500 for 30 min at 4 °C. Then, MSCs were washed twice with PBS again and analyzed by CytoFLEX S Flow cytometry (Beckman, USA).
For adipogenic and osteogenic differentiation, 2 × 10⁴ MSCs were seeded in a 6-well plate and maintained with osteogenic differentiation medium and adipogenic differentiation medium (Cyagen, USA) for 3 weeks. Mineralized nodule and lipid droplets were detected by alizarin red and Oil red O staining kit (Cyagen, USA), and observed under a fluorescence inverted microscope.

Xu H.J., Liu X.Z., Yang L., Ning Y., Xu L.L., Sun D.M., Liao W., Yang Y, & Li Z.H. (2023). Runx2 overexpression promotes bone repair of osteonecrosis of the femoral head (ONFH). Molecular Biology Reports, 50(6), 4769-4779.

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Protein Expression Analysis of Stem Cell Markers

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Total protein was extracted from cells and tissues with SDS lysis buffer (Beyotime), with the concentration measured utilizing a bicinchoninic acid kit (20201ES76, YEASEN Biotechnology Co., Ltd., Shanghai, China). After separation using 8% sodium dodecyl sulfate polyacrylamide gel electrophoresis, the sample was submitted to electrotransfer onto polyvinylidene fluoride membranes (Millipore, Billerica, MA, USA) which were blocked using 5% blocking solution with skimmed milk powder and underwent overnight incubation at 4°C with primary rabbit antibodies against CGRP (ab189786, 1 : 1000, Abcam), RUNX2 (ab76956, 1 : 1000, Abcam), OCN (ab93876, 1 : 500, Abcam), ALP (ab229126, 1 : 1000, Abcam), CD45 (ab40763, 1 : 5000, Abcam), CD73 (ab133582, 1 : 5000, Abcam), and CD90 (ab92574, 1 : 1000, Abcam) as well as 1 h-incubation with horseradish peroxidase-labeled secondary antibody goat anti-rabbit IgG (ab150077, 1: 1000, Abcam) at ambient temperature. The immunocomplexes on the membrane were visualized utilizing enhanced chemiluminescence (ECL) reagent (ECL808-25, Biomiga, USA) at ambient temperature for 1 min, and band intensities were quantified with the help of Image Pro Plus 6.0 software (Media Cybernetics, Silver Springs, MD, USA).

Yang Y., Zhang B., Yang Y., Peng B, & Ye R. (2022). PLGA Containing Human Adipose-Derived Stem Cell-Derived Extracellular Vesicles Accelerates the Repair of Alveolar Bone Defects via Transfer of CGRP. Oxidative Medicine and Cellular Longevity, 2022, 4815284.

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Characterization of hAMSCs by Immunofluorescence

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hAMSCs were fixed with 4% paraformaldehyde (Biosharp, China) for 20 min, and with permeabilized 0.1% Triton-X 100 (Sigma) for 15 min and were blocked with 5% bovine serum albumin (BSA) (Bosterbio) for 30 min at room temperature. The cells were incubated overnight at 4 °C with the primary antibodies anti-Integrin beta 1 (CD29) (ab134179, Abcam), anti-CD44 (ab189524, Abcam), anti-CD73 (ab133582, Abcam), anti-CD105 (ab231774, Abcam), anti-CD19 (ab134114, Abcam), anti-CD34 (ab81289, Abcam), anti-CD45 (ab40763, Abcam), anti-cytokeratin 7 (CK7) (ab181598, Abcam) and anti-cytokeratin 19 (CK19) (ab52625, Abcam) and then incubated with the Alexa Fluor 488-conjugated secondary antibody (ab150077, Abcam) for 1 h at 37 °C in the dark. The cell nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) (Sigma) for 5 min. The results were observed by fluorescence microscope.

Huang J., Zhang W., Yu J., Gou Y., Liu N., Wang T., Sun C., Wu B., Li C., Chen X., Mao Y., Zhang Y, & Wang J. (2022). Human amniotic mesenchymal stem cells combined with PPCNg facilitate injured endometrial regeneration. Stem Cell Research & Therapy, 13, 17.

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Immunophenotyping and Differentiation of hMSCs

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hMSCs were detached, centrifuged, and washed with phosphate-buffered saline (PBS), and then resuspended in the Stain Buffer and counted. The cell suspension was transferred to new Eppendorf tubes (1.5 mL), with about 5 × 10⁴ cells in each tube. According to the concentrations of antibodies recommended in the instructions of flow cytometry, 5 μL CD29 (ab263847, Abcam, Cambridge, UK), CD34 (ab81289, Abcam), CD45 (ab40763, Abcam), CD73 (ab202122, Abcam), CD90 (ab23894, Abcam), CD105 (ab2529, Abcam), CD117 (ab45924, Abcam), human leukocyte antigen-D-related (HLA-DR; ab92511, Abcam) antibodies and isotype controls were added to 50 μL cell suspension respectively. After being mixed evenly, the cell suspension containing antibodies was incubated for 30 min in a refrigerator at 4 °C in the dark, washed 3 times with pre-cooled Stain Buffer and centrifuged for 5 min at 300g. Then unbound antibodies were washed away. Finally, cells were resuspended in flow tubes with 500 μL Stain Buffer and detected by flow cytometry, and analyzed and processed using Flowjo 7.6 software.
OriCell^™ hMSC osteogenic differentiation kit, OriCell^™ hMSC adipogenic differentiation kit, and OriCell^™ hMSC chondrogenic differentiation kit were from Cyagen Biosciences (Guangzhou, China). The specific experiment operation was conducted following the instructions of the manufacturer’s kit.

Wang X., Jiang L, & Liu Q. (2022). miR-18a-5p derived from mesenchymal stem cells-extracellular vesicles inhibits ovarian cancer cell proliferation, migration, invasion, and chemotherapy resistance. Journal of Translational Medicine, 20, 258.

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Isolation and Characterization of Tendon-Derived Stem Cells

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TDSCs were harvested from Achilles tendon tissues of SD rats as reported previously [32 (link)]. Briefly, rat Achilles tendons were isolated and then incubated with collagenase (Sigma, Carlsbad, CA, USA) for 2 h at 37°C before harvesting TDSCs. Isolated cells were plated in 100 mm dishes at a density of 200 cells/cm² and cultured in DMEM containing 20% FBS (Gibco), 100 U/ml penicillin, and 100 U/ml streptomycin (Gibco) in a standard tissue culture incubator for 10 days. Cells were passaged up to three times before use. TDSCs were identified as reported previously [32 (link)]. Expression of stem cell surface markers on TDSCs was measured by flow cytometry with antibodies against CD45 (ab40763; Abcam, Cambridge, UK), CD34 (ab81289, Abcam), CD90 (ab33694, Abcam), CD44 (12-0444-80; eBioscience, Thermo Fisher Scientific, MA, USA), and CD160 (ab274374, Abcam). The percentages of positive cells were measured with the FACScan system (Becton Dickinson, San Jose, CA, USA).

Xue Z., Chen Z., Wu T., Li R., Chen C., Liu J., Hou H., Zheng X, & Wang H. (2022). VEGFA-Enriched Exosomes from Tendon-Derived Stem Cells Facilitate Tenocyte Differentiation, Migration, and Transition to a Fibroblastic Phenotype. BioMed Research International, 2022, 8537959.

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Immunostaining of Hematopoietic Cells

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The presence of infiltrating human hemopoietic CD45⁺ cells (monoclonal rabbit anti-human CD45 antibody, 1:500, #ab40763; Abcam, Cambridge, UK) and T cells (XP^® monoclonal rabbit anti-human CD3ε (D7A6E™), 1:300, #85061; Cell Signalling Technology^® Danvers, MA, USA) in murine tissue was detected by immunohistochemistry (secondary Goat-anti-Rabbit IgG H&L (HPR) antibody, 1:1000, #ab 205718; Abcam, Cambridge, UK) and immunofluorescence (secondary Goat-anti-Rabbit IgG H&L (Alexa Fluor^®) antibody, 1:6500, #111-585-003; Jackson Immunoresearch, West Grove, PA, USA) in Formalin-Fixed Paraffin-Embedded tissue blocks as previously described [16 (link),17 (link)].

Zeng B., Moi D., Tolley L., Molotkov N., Frazer I.H., Perry C., Dolcetti R., Mazzieri R, & Cruz J.L. (2023). Skin-Grafting and Dendritic Cell “Boosted” Humanized Mouse Models Allow the Pre-Clinical Evaluation of Therapeutic Cancer Vaccines. Cells, 12(16), 2094.

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CTC Enrichment and Identification from Whole Blood

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Whole blood samples were incubated with biotin-labeled CD45 antibody (Abcam ab40763, 1:100 dilution) and then exposed to streptavidin-labeled dynabeads to deplete white blood cells. Primary antibodies against EpCAM (Cell Signaling Technology VU1D9, 1:800) were added to the cell suspension for 1 h and then stained with Alexa Fluor® 488 (Abcam) to identify circulating tumor cells (CTCs). Enriched cells were washed with PBS and centrifuged at 800 g for 5 min. The sample was then detected using a BD FACSARIA III cell sorter (BD biosciences, USA).

Qiao Y., Liu C., Zhang X., Zhou Q., Li Y., Xu Y., Gao Z., Xu Y., Kong L., Yang A., Mei M., Ren Y., Wang X, & Zhou X. (2021). PD-L2 based immune signature confers poor prognosis in HNSCC. Oncoimmunology, 10(1), 1947569.

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Immune Profiling of Kidney Cancer

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After obtaining the patient’s consent and approval from the Ethics Committee of Qingdao Central Hospital (KY202318401), we obtained pathological sections from KIRC patients. Anti-LAG3 antibody (ab209236, 1:100 dilution), anti-CD45 antibody (ab40763, 1:100 dilution), and anti-PAX8 antibody (ab191870, 1:1000 dilution) were purchased from Abcam (Cambridge, UK). BSA (G5001), PBS (G0002), EDTA (G1203), and DAPI (G1012) were purchased from Servicebio (Wuhan, China). Paraffin-embedded tissue sections underwent xylene deparaffinization, followed by ethanol dehydration. Microwave antigen retrieval (EDTA buffer, pH 9.0, 10 minutes) was performed, and non-specific binding was blocked with BSA for 30 minutes. Anti-CD45, anti-PAX8, and anti-LAG3 antibodies were applied and incubated overnight at 4°C. After three PBS washes, sections were exposed to secondary antibodies for 50 minutes at room temperature under light protection. DAPI staining (10 minutes) for cell nuclei and a 5-minute application of autofluorescence quenching reagent were followed. Sections were then mounted on slides for microscopy, and images were captured using a fluorescence microscope.

Li J., Liu C., Su H., Dong H., Wang Z., Wang Y., Zhao P., Zhang C., Zhao Y, & Ma X. (2024). Integrative analysis of LAG3 immune signature and identification of a LAG3-related genes prognostic signature in kidney renal clear cell carcinoma. Aging (Albany NY), 16(3), 2161-2180.

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Multiplexed Imaging of Cancer Cells

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QDs-based multiplexed imaging was divided into two parts: QDs-based multiplexed imaging of cell climbing and QDs-based multiplexed imaging of substrate with captured cancer cells. For cell climbing staining, the cell climbing was treated with the following mixture of primary antibodies at 4°C overnight: mouse anti-human IgG monoclonal antibody against CK (dilution 1:600, BD-349205, BD Biosciences, Franklin Lakes, USA), goat anti-human IgG monoclonal antibody against vimentin (dilution 1:100, Ab-11256, Abcam, Cambridge, England) and a biotinylated anti-EpCAM monoclonal antibody. For the substrate staining, the substrate was incubated with the following mixture of primary antibodies at 4°C overnight: mouse anti-human IgG monoclonal antibody against CK, goat anti-human IgG monoclonal antibody against vimentin, and rabbit anti-human monoclonal antibody against CD45 (dilution 1:100, Ab-40763, Abcam). Then corresponding secondary antibodies, including QDs-585 goat F(ab')₂anti-mouse IgG conjugate (dilution 1:600, Q11011MP, Invitrogen, USA), QDs-655 rabbit F(ab')₂anti-goat IgG conjugate (dilution 1:600, Q11821MP, Invitrogen, USA), and QDs-labeled streptavidin-525 (dilution 1:100, QS525, Jiayuan Co., Ltd, China) or QDs-525 goat F(ab')₂ anti-rabbit IgG conjugate (dilution 1:50, Q11441MP, Invitrogen, USA) were added to the substrate and incubated at 37°C for 2 h (Fig. 1B1-B3).

Chen Y.Y., Cheng B.R., He Z.B., Wang S.Y., Wang Z.M., Sun M., Song H.B., Fang Y., Chen F.F, & Xiong B. (2016). Capture and Identification of Heterogeneous Circulating Tumor Cells Using Transparent Nanomaterials and Quantum Dots-Based Multiplexed Imaging. Journal of Cancer, 7(1), 69-79.

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Isolation and Analysis of Circulating Tumor Cells

Cited 4 times

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A total sample of 10 ml of blood from each cancer patient was collected in a Streck vacutainer tube and processed within 24 h after collection. CTCs were first enriched via mass-dependent microfluidics and then manually picked through immunofluorescent staining of both positive staining for pan-cytokeratins (panCK+) and negative staining for CD45 (CD45−) on the basis of an intact Hoechst-stained nucleus (blue fluorescence) as in the method previously reported (Hou et al., 2013 (link); Xu et al., 2018a (link); Abouleila et al., 2019 (link); Chemi et al., 2019 (link)). Anti-CD45 (AB40763, Abcam, United States), anti-panCK (AB7753, Abcam, United States), and Hoechst 33342 were used for imaging of CTCs and white blood cells (WBCs) (Figure 1), which were counted under a fluorescence microscope (Olympus IX73), manually picked with a glass capillary into an ice-precooling low-adsorption PCR tube, and stored at −80°C for further experiments. To prevent sequencing failures, biological triplicates to quintuplicates of 10 CTCs were collected from each patient. Biological triplicates of 10 WBCs were collected from the blood sample of P8 and used as the reference control for calling mutations in 10-CTC WES data sets after whole genome amplification (WGA).

Chang Y., Wang Y., Li B., Lu X., Wang R., Li H., Yan B., Gu A., Wang W., Huang A., Wu S, & Li R. (2021). Whole-Exome Sequencing on Circulating Tumor Cells Explores Platinum-Drug Resistance Mutations in Advanced Non-small Cell Lung Cancer. Frontiers in Genetics, 12, 722078.

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