Ab150080

Manufactured by Abcam

Sourced in United Kingdom, United States, Germany, China

Ab150080 is a primary antibody that can be used for immunohistochemistry applications. It is a rabbit polyclonal antibody.

Automatically generated - may contain errors

Lab products found in correlation

Ab150113, by Abcam (45 mentions) Ab150077, by Abcam (36 mentions) Ab150116, by Abcam (15 mentions) Ab104139, by Abcam (12 mentions) Ab150117, by Abcam (7 mentions) Hoechst 33342, by Thermo Fisher Scientific (7 mentions) Lsm 710, by Zeiss (6 mentions) Alexa fluor 488, by Abcam (5 mentions) Ab28364, by Abcam (5 mentions) Ab15580, by Abcam (5 mentions) Ab150115, by Abcam (5 mentions) Alexa fluor 594, by Abcam (4 mentions) Fluorescence microscope, by Olympus (4 mentions) Ab150105, by Abcam (4 mentions) Ab125212, by Abcam (4 mentions) C0065, by Solarbio (4 mentions) Cm1950, by Leica (4 mentions) Fluorescence microscope, by Leica (4 mentions) Lsm 800, by Zeiss (4 mentions) Lsm 880, by Zeiss (4 mentions)

Close spelling variants of this product

Alexa fluor-594 conjugated anti-rabbit secondary antibody (Cat no. ab150080) (2 citations) Ab150080/ab150116 (2 citations)

207 protocols using ab150080

Immunofluorescence Analysis of Macrophage Markers

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Colon sections were selected after deparaffinization was performed with 10 mM, pH 6.0 sodium citrate solution in 95°C water bath for 10 min, followed by a 20 min incubation at room temperature. Slides were washed, blocked in 5% normal goat serum at 37°C, and stained using the primary antibodies that rabbit anti-F4/80 antibody (29414-1-AP, Proteintech) and mouse anti-CD206 antibody (60143-1-Ig, Proteintech) overnight at 4°C, respectively. Then sections were incubated with goat anti-rabbit alexa fluor 594 (ab150080, Abcam) and goat anti-mouse alexa fluor 488 (A0428, Beyotime) for 1 hour. The sections were photographed with a confocal microscope (FLUOVIEW FV3000, Olympus) and analyzed with Image J software (National Institutes of Health, Bethesda, MD).
Cell samples were fixed for 30 min by 4% paraformaldehyde, permeated for 10 min in 0.03% Triton-X-100. Then, 5% goat serum was incubated for 30 min at 37°C, and incubated in METTL3 antibody (15073-1-AP, Proteintech) overnight at 4°C. The next day, cells were incubated with goat anti-rabbit alexa fluor 594 (ab150080, Abcam) for 1 hour, DAPI stained the nuclei for 10 min, and then photographed.

Li Y., Ma B., Wang Z., Chen Y, & Dong Y. (2024). The Effect Mechanism of N6-adenosine Methylation (m6A) in Melatonin Regulated LPS-induced Colon Inflammation. International Journal of Biological Sciences, 20(7), 2491-2506.

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Astrocyte and Neuron Characterization

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The isolated astrocytes or neurons were characterized by staining marker GFAP or Neun. Firstly, cells were put in 4% paraformaldehyde (Biosharp, Beijing, China) for 20 min. After that, put them in PBS containing 0.1% Triton X100 (Beyotime, Shanghai, China) for 20 min. Then incubated them with 5 % BSA (Biofroxx, Shanghai, China) for 1 h. After above, primary astrocytes were incubated with rabbit monoclonal antibody anti-GFAP (1:200; #80788S; CST, USA) and primary neurons were incubated with a rabbit monoclonal antibody anti-NeuN (1:200; #24307; CST) overnight at 4° C and then incubated the next day with goat anti-rabbit IgG H&L (Alexa Fluor® 488) (1:200; ab150077; Abcam, Cambridge, MA, USA) or goat anti-rabbit IgG H&L (Alexa Fluor® 594) (1:200; ab150080; Abcam) and incubated with DAPI (1:300, Cat. # C1002, Beyotime) to stain nuclei. Finally, the images were observed and photographed. The number of GFAP-positive cells or Neun-positive cells was calculated from randomly selected microscopic fields. Astrocyte or neuronal purity was presented as percentage of the number of the percentage of GFAP-positive or Neun-positive cells to the total cells of view, both around 90% or more.

Hu J., Duan H., Zou J., Ding W., Wei Z., Peng Q., Li Z., Duan R., Sun J, & Zhu J. (2024). METTL3-dependent N6-methyladenosine modification is involved in berberine-mediated neuroprotection in ischemic stroke by enhancing the stability of NEAT1 in astrocytes. Aging (Albany NY), 16(1), 299-321.

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Immunofluorescence Analysis of Neuronal Markers

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For immunofluorescence analysis, cell cultures were washed twice with PBS, fixed with 4% paraformaldehyde for 15 min, washed three additional times with PBS, permeabilized with 0.1% Triton X-100 in PBS for 5 min at 37 °C, and blocked for 2 h with 4% albumin. The cells were then incubated overnight with either anti-beta III tubulin (1:400, ab78078, Abcam), anti–HIF–1a (NB100105, Novus), or anti-GFAP (#80788, CST) antibodies at 4 °C, washed with PBS, and incubated with a goat anti-mouse secondary antibody conjugated with either a goat anti-rabbit secondary antibody (Alexa Fluor 488, ab150077 Abcam), goat anti-mouse secondary antibody (Alexa Fluor 488, ab150113, Abcam) or goat anti-rabbit secondary antibody (Alexa Fluor 594, ab150080, Abcam) for 1 h at room temperature. Cell nuclei were stained with 0.2 μg/mL of 4,6-diamino-2-phenylindole (DAPI, D9542, Sigma–Aldrich).
ImageJ software with the Sholl analysis plugin was used to quantify the neurite length, the number of neurites from soma, the number of secondary branches, and the number of neurite intersections according to a previous protocol [25 (link)]. The analyzers were blinded to the neuron groups.

Zheng J., Xie Y., Ren L., Qi L., Wu L., Pan X., Zhou J., Chen Z, & Liu L. (2021). GLP-1 improves the supportive ability of astrocytes to neurons by promoting aerobic glycolysis in Alzheimer's disease. Molecular Metabolism, 47, 101180.

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Pentobarbital Sodium Protocol for Cell Culture

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Pentobarbital sodium was purchased from Shanghai Rongbai Biological Technology Co. Ltd. DMEM (11,995,073), fetal bovine serum (FBS, 10,100,147), phosphate buffered saline (PBS) and penicillin-streptomycin (15,140,163) were obtained from Gibco (Carlsbad, CA, USA). Rabbit monoclonal anti-rat alpha-smooth muscle actin (α-SMA, 1:500, ab124964) and goat polyclonal anti-rabbit IgG-H&L (1:1000, ab150080) antibodies were purchased from Abcam. Other experimental materials were purchased from Thermo.

Huang W., Liu H., Pan Y., Wang X., Yang H., Wang D., Lin J, & Zhang H. (2023). A modified primary culture method of rat pulmonary vein smooth muscle cells. Journal of Cardiothoracic Surgery, 18, 146.

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NF-κB Nuclear Translocation Inhibition by 5-HMF

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NF-κB nuclear translocation was examined by immunofluorescence microscopy. RAW 264.7 cells (5 × 10⁴ cells/each well in a 24-well plate with sterile coverslips) were pre-treated with 0, 31.5, 63.0 and 126.0 μg/mL of 5-HMF for 6 h and then stimulated by 1 μg/mL of LPS for 18 h. Cells were fixed with 4% paraformaldehyde for 15 min at room temperature and then washed with PBS twice. Later, the cells were washed with a quenching solution (0.1% glycine in PBS) twice, permeated with 0.1% Triton X-100 for 10 min and blocked using a blocking solution (10% FBS in PBS) for 1 h at room temperature. Then the cells were incubated with the primary p65 antibody at 4 °C overnight and later incubated with Conjugated Goat anti-Rabbit IgG H&L (Alexa Fluor^® 594) (1:2000, ab150080, Abcam, UK) for 1 h at room temperature and then stained with 4,6-diamidino-2 phenylindole (DAPI) for 5 min. The coverslips were removed and observed using laser scanning confocal microscope (Nikon Ti-E A1, Japan).

Kong F., Lee B.H, & Wei K. (2019). 5-Hydroxymethylfurfural Mitigates Lipopolysaccharide-Stimulated Inflammation via Suppression of MAPK, NF-κB and mTOR Activation in RAW 264.7 Cells. Molecules, 24(2), 275.

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Visualizing Vascular Tissue Formation

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We performed immunofluorescence staining of F-actin and VE-cadherin expressed by the microengineered endothelium to demonstrate the formation of physiological vascular tissue in our model. As the first step of this procedure, the cells were fixed with a 4% paraformaldehyde solution for 15 min, and the reversibly bonded microchannel layer was removed from the microdevice to gain direct access to the cells on the bottom channel surface. The cells were then permeabilized with 0.5% Triton X-100 for 15 min. After washing, the cells were incubated with Alexa Fluor 488 conjugated Phalloidin (A12379, Life Technologies, Carlsbad, CA) and DAPI for 20 min to stain F-actin and nuclei, respectively. For visualization of adherens junctions, the cells were first incubated overnight with primary VE-cadherin antibody (2500 S, Cell Signaling, Danvers, MA) diluted in 2% BSA solution. Secondary anti-rabbit antibody conjugated to Alexa Fluor 594 (ab150080, Abcam, Cambridge, MA) was diluted in 2% BSA solution and incubated for 1 hour. Nuclei were labeled with DAPI. Images of the fluorescently stained cells were acquired by confocal microscopy (Leica, Wetzlar, Germany).

Seo J., Conegliano D., Farrell M., Cho M., Ding X., Seykora T., Qing D., Mangalmurti N.S, & Huh D. (2017). A microengineered model of RBC transfusion-induced pulmonary vascular injury. Scientific Reports, 7, 3413.

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Molecular Profiling of Neuropathic Pain

Cited 1 time

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After completing the last behavioral tests at 48 h, all rats were sacrificed
under deep anesthesia (5% sevoflurane). According to the literature,^{27 (link)} the L3-L5
right DRGs were rapidly removed and stored in liquid nitrogen. Tissue samples
were homogenized in lysis buffer, which was centrifuged at 12,000 r/min for
10 min at 4°C to collect the supernatant. The protein concentration was measured
by BCA Protein Assay Kit (Solarbio, Beijing, China). Protein samples were
separated on SDS-polyacrylamide gel electrophoresis, and the fractionated
proteins were transferred to a polyvinylidene difluoride (PVDF) membrane (Thermo
Fisher Scientific, USA). After being blocked with 5% (w/v) skimmed milk for 1 h
at room temperature, the membranes were incubated overnight at 4°C with the
diluted primary antibodies. The primary antibodies used were: anti-TRPA1
(GTX54765, GeneTex, USA), anti-TLR4 (GTX57153, GeneTex, USA), anti- NF-κB
(ab194726, Abcam, USA), and anti-GAPDH (ab8245, Abcam, USA). The membrane was
washed with Tris-buffered saline with Tween buffer and incubated with the goat
anti-mouse secondary antibody (ab150113, Abcam, USA) or goat anti-rabbit
secondary antibody (ab150080 Abcam, USA) for 1 h at room temperature. The bands
were scanned by ECL Western Blotting Substrate (Tanon, China) and analyzed by
Image J software.

Liu X., Gong R., Peng L, & Zhao J. (2023). Toll-like receptor 4 signaling pathway in sensory neurons mediates remifentanil-induced postoperative hyperalgesia via transient receptor potential ankyrin 1. Molecular Pain, 19, 17448069231158290.

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Immunofluorescence Analysis of Stem Cell Markers

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BMSCs were washed with precooled PBS, followed by fixation with 4% paraformaldehyde for 10 min as well as permeability with 0.25% Triton-X100 for 15 min at room temperature. Afterwards, blockage with 1% bovine serum albumin was implemented for 60 min, along with incubation with primary antibody of LIF (1 : 100; ab113262, Abcam), LIFR (1 : 100; ab232877), alkaline phosphatase (ALP; 1 : 200; ab224335), bone sialoprotein (BSP; 1 : 200; ab270605), and osteocalcin (OCN; 1 : 200; ab198228) at 4°C overnight as well as Alexa Fluor® 488- (1 : 200; ab150077; Abcam) or Alexa Fluor® 594- (1 : 200; ab150080; Abcam) conjugated secondary antibody for 1 h. Thereafter, BMSCs were stained with DAPI for 15 min. Images were captured utilizing a fluorescence microscope (Olympus, Japan).

Liang Y., Zhou R., Liu X., You L., Chen C., Ye X., Wei W., Liu J., Dai J., Li K, & Zhao X. (2022). Leukemia Inhibitory Factor Facilitates Self-Renewal and Differentiation and Attenuates Oxidative Stress of BMSCs by Activating PI3K/AKT Signaling. Oxidative Medicine and Cellular Longevity, 2022, 5772509.

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Quantification of Autophagic Activity in VSMCs

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The VSMCs were cultured in a culture dish with cover glasses placed on top. When cell confluence reached 50%, the cover glass was removed. The cells were then rinsed three times using PBS and fixed in 4% paraformaldehyde for 30 minutes at room temperature. After 15 minutes of permeation using 2% Triton X‐100, the cells were sealed for 45 minutes using 2% BSA. The sealing solution was then discarded, whereupon the cells were subjected to overnight incubation at 4°C with LC3 II antibody (ab63817, 1:100, Abcam Inc). After three PBS washes, the cells were re‐probed with secondary goat anti‐rabbit IgG H&L (ab150080, 1:400, Abcam Inc) at room temperature for 2 hours. Then 4′, 6‐diamidino‐2‐phenylindole DAPI (2 μg/mL) was added for cell staining followed by mounting on glass slides. The expression of LC3 II was then detected under a fluorescence microscope, and the ImageJ software was used to quantify the fluorescence intensity.

Zhou J., Zhou H., Liu C., Huang L., Lu D, & Gao C. (2020). HDAC1‐mediated deacetylation of LSD1 regulates vascular calcification by promoting autophagy in chronic renal failure. Journal of Cellular and Molecular Medicine, 24(15), 8636-8649.

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Immunofluorescence Analysis of Arterial Proteins

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Artery samples were fixed in 4% paraformaldehyde solution for paraffin embedding. Frozen tissues were embedded in the O.C.T. compound (Tissue-Tek). Sections were blocked in 5% goat serum for 1 h at room temperature, then incubated with primary antibody at 4 °C overnight. The primary antibodies used in this study was as follows. CD31 (1:100, #ab28364, Abcam; 1:100, #ab9498, Abcam), caveolin-1 (1:50, #3267, Cell Signaling Technology), Sp1 (1:50, #sc−420, Santa Cruz Biotechnology Inc.), Sp3 (1:50, #sc-28305, Santa Cruz Biotechnology Inc.), p-AMPKα (Thr172, 1:100, #50071, Cell Signaling Technology). After washing with phosphate buffered saline, samples were incubated with Alexa Fluor-labeled secondary antibody at room temperature for 1 h including Alexa Fluor 488 (1:200, #ab150077, Abcam; 1:200, #ab150113, Abcam), Alexa Fluor 594 (1:200, #ab150080, Abcam; 1:200, #ab150116, Abcam). Finally, nuclei were stained with 4,6’-diamidino-2-phenylindole (DAPI, #ab104139, Abcam) for 5 min at room temperature, and immunofluorescence was analyzed under a florescent microscope.

Lu H., Jiang X., He L., Ji X., Li X., Liu S., Sun Y., Qin X., Xiong X., Philipsen S., Xi B., Zhang M., Yang J., Zhang C., Zhang Y, & Zhang W. (2023). Endothelial Sp1/Sp3 are essential to the effect of captopril on blood pressure in male mice. Nature Communications, 14, 5891.

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