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Ab75769

Manufactured by Abcam
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Ab75769 is a lab equipment product manufactured by Abcam. It is designed to perform a core function related to laboratory research, but a detailed description of its intended use cannot be provided in a concise, unbiased, and factual manner.

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Lab products found in correlation

Ab11414, by Abcam (2 mentions) Ab32570, by Abcam (2 mentions) Ab109536, by Abcam (2 mentions) Ab13979, by Abcam (2 mentions) Ab11306, by Abcam (2 mentions) Ab74524, by Abcam (2 mentions) Ab96569, by Abcam (2 mentions) Ab76956, by Abcam (2 mentions) Ab108337, by Abcam (2 mentions) Ab34710, by Merck Group (2 mentions) Ab33734, by Abcam (2 mentions) Ab189267, by Abcam (2 mentions) Ab203887, by Abcam (2 mentions) Ab185716, by Abcam (2 mentions) Ab107595, by Abcam (2 mentions) Ab93876, by Abcam (2 mentions) Ab19027, by Abcam (2 mentions) Mab8887, by Abcam (2 mentions) Ab16667, by Abcam (2 mentions) Ab81289, by Abcam (2 mentions)

36 protocols using ab75769

1

Immunohistochemical Analysis of Periodontal Tissue

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The trimmed maxillae were fixed in 10% neutral buffered formalin for 24 h, decalcified in 15% ethylenediaminetetraacetic acid and embedded in paraffin. Consecutive 4-μm sagittal sections of the maxillary first molar were obtained and the middle to apical third of the periodontal tissues was observed. A two-step detection kit (Zhongshan Golden Bridge Biotechnology, Beijing, China) was used following previous protocols (He et al., 2015b (link)). Primary antibodies including anti-microtubule associated protein 1 light chain 3 β (LC3B) (1:100, CST3868S, CST), anti-TNF-α (1:100, ab1793, Abcam), and anti-CD146 (1:200, ab-75769, Abcam) were used. Five different slides from each sample (n = 10) were used for cell counting. Each slide was measured for three times and the average positive cell numbers were calculated.
Jiang N., He D., Ma Y., Su J., Wu X., Cui S., Li Z., Zhou Y., Yu H, & Liu Y. (2021). Force-Induced Autophagy in Periodontal Ligament Stem Cells Modulates M1 Macrophage Polarization via AKT Signaling. Frontiers in Cell and Developmental Biology, 9, 666631.
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2

Chondrocyte Immunophenotyping by Flow Cytometry

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Chondrocytes (1×106) from growth plate (passage 1) were washed in PBS and incubated with corresponding primary antibodies for one hour at 4°C. Primary antibodies used here were conjugated antibodies specific for CD44-FITC (dilution 1:100, #550974; BD Biosciences, San Diego, CA, USA), CD29-PE (dilution 1:80, #48-0291; eBiosciences; Thermo Fisher Scientific, Inc.), CD34-PE (dilution 1:100; #sc-74499 PE; Santa Cruz Biotechnology, Inc., Dallas, TX, USA), CD45-PE (dilution 1:80, #12-0461; eBiosciences; Thermo Fisher Scientific, Inc.) and unconjugated antibodies for CD105 (dilution 1:100, #ab11414) and CD146 (dilution 1:80, #ab75769) (both from Abcam, Cambridge, MA, USA). After washing with PBS, cells stained with CD105 and CD146 were incubated with anti-mouse FITC-conjugated secondary antibody (dilution 1:500, #A0568; Beyotime Institute of Biotechnology, Guangzhou, China) or Alexa Fluor 594 donkey anti-rabbit IgG (dilution 1:500, #A-21207; Invitrogen; Life Technologies) for 30 min at 4°C. The cells were washed twice and then re-suspended in 200 µl PBS for the flow cytometry analysis (FACSort; BD Biosciences).
Wu Y.X., Jing X.Z., Sun Y., Ye Y.P., Guo J.C., Huang J.M., Xiang W., Zhang J.M, & Guo F.J. (2017). CD146+ skeletal stem cells from growth plate exhibit specific chondrogenic differentiation capacity in vitro. Molecular Medicine Reports, 16(6), 8019-8028.
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3

Immunofluorescence Analysis of Vascular Markers

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UCA, UCV, and WJ were immersed in optimum cutting temperature (OCT) compound (Leica, Wetzlar, Germany) and frozen at −70°C until sectioning. The tissues were serially sectioned to 6 μm thickness using a cryostat (Leica). Expression of PDGF-Rβ (ab32570, Abcam, Cambridge, UK), NG2 (ab139406, Abcam), α-SMA (ab5694, Abcam), and CD146 (ab75769, Abcam) was detected by immunofluorescence staining. After incubated with primary antibody at 4°C overnight, the frozen sections were then incubated with Alexa Fluor 488-conjugated goat anti-rabbit IgG (1 : 200, Invitrogen, Grand Island, NY, USA) or Alexa Fluor 555-conjugated goat anti-rabbit IgG (1 : 200, Invitrogen). The nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI), which was contained in the Vectashield mounting medium for fluorescence (Vector Laboratories Inc.). The images were visualized using fluorescence confocal microscopy (Leica) under a magnification of 600x. The integrated optical density (IOD) values of positive staining in five randomly selected fields of view were tested by Image pro-plus 6.0 software (Media Cybernetic, Rockville, USA).
Xu L., Zhou J., Liu J., Liu Y., Wang L., Jiang R., Diao Z., Yan G., Pèault B., Sun H, & Ding L. (2017). Different Angiogenic Potentials of Mesenchymal Stem Cells Derived from Umbilical Artery, Umbilical Vein, and Wharton's Jelly. Stem Cells International, 2017, 3175748.
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4

3D Visualization of Cell Response in Tendons

Cited 2 times
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To enable cell response to injury to be visualised in 3D, PFA-fixed tendons (n = 2 per time point) were immunolabelled and optically cleared using HISTO™ solutions (Visikol, Hampton, NJ, USA) according to our recently developed protocol [11 (link)]. Briefly, following permeabilisation and blocking, tendons were incubated with primary antibodies for CD146 (1:100; Abcam: ab75769, Cambridge, UK) for 96 h to allow for the complete diffusion of the antibodies through the full thickness of the tendon. Secondary antibody incubation (Alexa Fluor® 594 Goat anti-rabbit IgG, Invitrogen, 1:500, Waltham, MA, USA) was performed for 96 h, followed by overnight nuclei counterstaining with DAPI. Some tendons were additionally stained with Col-F (20 µm; ImmunoChemistry Technologies, Bloomington, MN, USA), a fluorescent probe that binds collagen and elastin, during the nuclei counterstaining step to allow for the visualisation of collagen within the tendon in 3D [87 (link)]. Tendons were then washed, dehydrated in methanol and cleared by incubation in HISTO™-1 and -2 solutions for at least 72 h. Tendons were stored in HISTO™-2 prior to imaging.
Marr N., Meeson R., Kelly E.F., Fang Y., Peffers M.J., Pitsillides A.A., Dudhia J, & Thorpe C.T. (2021). CD146 Delineates an Interfascicular Cell Sub-Population in Tendon That Is Recruited during Injury through Its Ligand Laminin-α4. International Journal of Molecular Sciences, 22(18), 9729.
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5

Western Blot Analysis of CD146

Cited 1 time
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Western blot assays were used according to standard techniques as previously reported (40 (link)). Antibodies against CD146 (ab75769; Abcam) and β-actin (#3700; CST) were used.
Lv Z., Feng H.Y., Tao W., Li H.Z, & Zhang X. (2021). CD146 as a Prognostic-Related Biomarker in ccRCC Correlating With Immune Infiltrates. Frontiers in Oncology, 11, 744107.
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6

Immunohistochemical Analysis of PDX Tissues

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The paraffin embedded PDX tissues sections (5μm) on glass slides were deparaffinized and hydrated, and antigen retrieval was performed using a decloaker with a target retrieval solution (pH, 6.0; Dako). The intrinsic peroxidase activity was blocked using 3% methanol and hydrogen peroxide for 10 minutes, and a serum-free protein block (Dako) was used for 5 minutes to block nonspecific antibody binding. The slides were then incubated with antibodies against human MCAM (ab75769, 1:200 dilution; Abcam), EGFR (ab52894, 1:100, Abcam), EPHA2 (#6997, 1:200, Cell Signaling), ITGB1 (ab52971, 1:250, Abcam) and JAG1 (ab109536, 1:100, Abcam) overnight at 4°C. After being washed three times in Tris-buffered saline, the slides were then incubated for 30 minutes with Dako EnVision+ Dual Link at room temperature. Slides were incubated with Dako chromogen substrate for 5 minutes and counterstained with hematoxylin. Formalin-fixed, paraffin-embedded, whole-section specimens with the primary antibodies omitted were used as negative controls.
Tripathi S.C., Fahrmann J.F., Celiktas M., Aguilar M., Marini K.D., Jolly M.K., Katayama H., Wang H., Murage E.N., Dennison J.B., Watkins D.N., Levine H., Ostrin E.J., Taguchi A, & Hanash S.M. (2017). A Novel Mechanism of Chemoresistance in Small Cell Lung Cancer mediated by MCAM via PI3K/AKT/SOX2 Signaling Pathway. Cancer research, 77(16), 4414-4425.
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7

Characterization of hJBMMSCs

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Cultured hJBMMSCs were isolated and stained with primary CD29 mAb (303005, Biolegend), CD13 mAb (ab7417, Abcam), CD45 mAb (D9M8I, Cell Signaling Technology, Beverly, MA), and CD146 mAb (ab75769, Abcam), which was followed by donkey anti-rabbit FITC/CY3 conjugated secondary mAb (Life Technologies). hJBMMSCs were then analyzed on a FACSCalibur cytometer (BD Biosciences) and identified as CD29+CD13+CD45−CD146−.
Chen X., Liu Y., Ding W., Shi J., Li S., Liu Y., Wu M, & Wang H. (2018). Mechanical stretch-induced osteogenic differentiation of human jaw bone marrow mesenchymal stem cells (hJBMMSCs) via inhibition of the NF-κB pathway. Cell Death & Disease, 9(2), 207.
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8

Immunohistochemical Analysis of FABP4 and CD146

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Sections were deparaffinized, rehydrated in graded ethanol, boiled in citrate buffer for antigen retrieval, and stained with a primary antibody for FABP4 (1:200, #ab13979, Abcam, Cambridge, UK) and CD146 (1:200, #ab75769, Abcam). HRP-conjugated secondary antibody (#87–8963, Invitrogen) was used (#00–1111, Invitrogen).
Hu L., Yang G., Hägg D., Sun G., Ahn J.M., Jiang N., Ricupero C.L., Wu J., Rodhe C.H., Ascherman J.A., Chen L, & Mao J.J. (2015). IGF1 Promotes Adipogenesis by a Lineage Bias of Endogenous Adipose Stem/Progenitor Cells. Stem cells (Dayton, Ohio), 33(8), 2483-2495.
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9

Characterization of Musculoskeletal Cell Markers

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Primary antibodies used were specific for Collagen type I (Abcam, ab34710, 1:100 dilution), Collagen type II (Millipore, MAB8887, 1:100 dilution), murine CD200 (Abcam, ab33734, 1:100 dilution), Nestin (Abcam, ab11306, 1:200 dilution), Gremlin 1 (Abcam, ab189267, 1:50 dilution), COMP (Abcam, ab74524, 1:50 dilution), Aggrecan (Abcam, ab3778, 1:100 dilution), Thy1.2 (Invitrogen, cat no. 14-0902-82, 1:50 dilution, 6C3 (Invitrogen, cat no. 14-5891-82 (1:50 dilution), CD105 (Abcam, ab107595, 1:100 dilution), Runx2 (Abcam, ab76956, 1:200 dilution), Alpl (Abcam, ab108337, 1:100 dilution), Osteocalcin (Abcam, ab93876, 1:100 dilution), CD146 (Abcam, ab75769, 1:100 dilution), CD140α (Abcam, ab96569, 1:100 dilution), CD200 (Abcam, ab203887, 1:200 dilution), Tartrate Resistant Acid Phosphatase (TRAP) (Abcam, ab185716, 1:50 dilution), and Cathepsin K (Abcam, ab19027, 1:200 dilution).
Debnath S., Yallowitz A.R., McCormick J., Lalani S., Zhang T., Xu R., Li N., Liu Y., Yang Y.S., Eiseman M., Shim J.H., Hameed M., Healey J.H., Bostrom M.P., Landau D.A, & Greenblatt M.B. (2018). Discovery of a periosteal stem cell mediating intramembranous bone formation. Nature, 562(7725), 133-139.
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10

Characterization of Musculoskeletal Cell Markers

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Primary antibodies used were specific for Collagen type I (Abcam, ab34710, 1:100 dilution), Collagen type II (Millipore, MAB8887, 1:100 dilution), murine CD200 (Abcam, ab33734, 1:100 dilution), Nestin (Abcam, ab11306, 1:200 dilution), Gremlin 1 (Abcam, ab189267, 1:50 dilution), COMP (Abcam, ab74524, 1:50 dilution), Aggrecan (Abcam, ab3778, 1:100 dilution), Thy1.2 (Invitrogen, cat no. 14-0902-82, 1:50 dilution, 6C3 (Invitrogen, cat no. 14-5891-82 (1:50 dilution), CD105 (Abcam, ab107595, 1:100 dilution), Runx2 (Abcam, ab76956, 1:200 dilution), Alpl (Abcam, ab108337, 1:100 dilution), Osteocalcin (Abcam, ab93876, 1:100 dilution), CD146 (Abcam, ab75769, 1:100 dilution), CD140α (Abcam, ab96569, 1:100 dilution), CD200 (Abcam, ab203887, 1:200 dilution), Tartrate Resistant Acid Phosphatase (TRAP) (Abcam, ab185716, 1:50 dilution), and Cathepsin K (Abcam, ab19027, 1:200 dilution).
Debnath S., Yallowitz A.R., McCormick J., Lalani S., Zhang T., Xu R., Li N., Liu Y., Yang Y.S., Eiseman M., Shim J.H., Hameed M., Healey J.H., Bostrom M.P., Landau D.A, & Greenblatt M.B. (2018). Discovery of a periosteal stem cell mediating intramembranous bone formation. Nature, 562(7725), 133-139.
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