The cells were fixed with 4% formaldehyde for 30 min at 4°C and then permeabilized with 0.1% Triton X-100 in PBS for 15 min at 4°C. After blocking with 10% FBS for 30 min at 37°C, the cells were incubated with anti-IBA (cat no. ab178846; 1:200) and anti-AT1R primary antibodies (cat no. ab124505; 1:200) (both from Abcam) at 4°C overnight. Subsequently, the cells were incubated with goat anti-rabbit IgG H&L Alexa Fluor® 488 antibody (cat no. ab150077; 1:1,000; Abcam) for 30 min at 37°C. The nuclei were stained with DAPI after IF staining for 15 min at room temperature. Images were visualized under a fluorescence microscope (Eclipse 80i; Nikon Corporation).
Goat anti rabbit igg h l alexa fluor 488 antibody
Manufactured by Abcam
Sourced in United Kingdom
Goat anti-rabbit IgG H&L (Alexa Fluor® 488) antibody is a secondary antibody conjugated with Alexa Fluor® 488 dye. It is designed to detect and bind to rabbit immunoglobulin G (IgG) heavy and light chains.
Automatically generated - may contain errors
Lab products found in correlation
Eclipse 80i, by Nikon (1 mentions)
Fmoc amino acids, by GL Biochem (1 mentions)
Alkaline phosphatase alp, by Yeasen (1 mentions)
β alanine, by Solarbio (1 mentions)
Anti acetyl histone h3, by Merck Group (1 mentions)
Parp rabbit mab, by Cell Signaling Technology (1 mentions)
Anti acetyl histone h4 lys8, by Merck Group (1 mentions)
Fitc annexin 5 apoptosis detection kit, by BD (1 mentions)
2 chlorotrityl chloride resin, by GL Biochem (1 mentions)
Lysotracker red, by Yeasen (1 mentions)
β actin mouse mab, by Proteintech (1 mentions)
Paxillin antibody, by Abcam (1 mentions)
Confocal laser scanning microscope, by Zeiss (1 mentions)
Ptbp1 antibody, by Thermo Fisher Scientific (1 mentions)
Goat anti mouse igg h l alexa fluor 647 antibody, by Abcam (1 mentions)
Hoechst 33342, by Beyotime (1 mentions)
Confocal microscopy, by Leica (1 mentions)
Cytation 5 cell imaging multi mode reader, by Agilent Technologies (1 mentions)
Goat anti mouse igg h l alexa fluor 594 antibody, by Abcam (1 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (1 mentions)
8 protocols using goat anti rabbit igg h l alexa fluor 488 antibody
1
Immunofluorescence Staining of IBA-1 and AT1R
2
Protein Expression and Apoptosis Analysis
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Fmoc-amino acids and 2-chlorotrityl chloride resin were purchased from GL Biochem (Shanghai, China). β-Alanine, 4-chloro-7-nitro-1,2,3-benzoxadiazole (NBD-Cl), crystal violet, bovine serum albumin (BSA), DAPI, propidium iodide (PI), l -phenylalanine and other chemical reagents were bought from Solarbio Science & Technology Co., Ltd. (Beijing, China). Alkaline phosphatase (ALP) and Lyso-Tracker Red were obtained from Yeasen Biotechnology (Shanghai, China). Dulbecco's modified Eagle's medium (DMEM), RPMI Medium 1640 basic and fetal bovine serum (FBS) were obtained from Gibco (Suzhou, China). Anti-γH2AX (phosphor A139) antibody and goat anti-rabbit IgG H&L (Alexa Fluor® 488) antibody were obtained from Abcam (Shanghai, China). PARP rabbit mAb, cleaved PARP (Asp214) rabbit mAb and caspase 3 rabbit antibody were purchased from Cell Signaling Technology (Shanghai, China). Anti-acetyl histone H3 and anti-acetyl histone H4 (Lys 8) antibodies were provided by Millipore (Billerica, USA). β-Actin Mouse mAb and peroxidase-conjugated affinipure goat anti-mouse/rabbit IgG (H + L) were bought from Proteintech Group Inc. (Wuhan, China). FITC Annexin V apoptosis detection kit was purchased from BD Pharmingen (San Diego, CA, USA). Dynasore was purchased from TCI Development Co., Ltd. (Shanghai, China).
3
Immunofluorescence Staining of Paxillin and PTBP1
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Grow cells in 6-well plates containing confocal slides (1 × 104 cells per well). After washing with PBS, cells were fixed with 4% paraformaldehyde for 15 min, and 0.5%Triton X-100 was permeable for 20 min. After the slides were soaked with PBS, 1%bovine serum albumin was blocked for 30 min, each slides were dripped with sufficient amount of diluted Paxillin antibody (Abcam, UK, Cat. no. ab32084) and incubated overnight at 4℃ in a wet box. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody (Abcam, UK, Cat. no. ab150077) was added in the dark and incubated in a wet box at 37℃. Then, cells were incubated overnight at 4℃ in a wet box with diluted PTBP1 antibody (Invitrogen, USA, Cat. no. 32-4800), followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) antibody (Abcam, UK, Cat. no. ab150115). Hoechst 33342 (Beyotime Biotechnology, China, Cat. no. C1025) was added to the drops and incubated for 5 min, and the specimens were nucleated. The tablets were sealed with a sealing solution containing an anti-fluorescence quench agent, and the acquired images were observed under a confocal laser scanning microscope (Carl Zeiss, Germany) at 200x magnification.
4
Immunofluorescence Staining of Cultured Cells
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Cells cultured on coverslips were washed thrice by cold phosphate-buffered saline (PBS) for and then fixed in 4% formaldehyde for 15 min. Then we added 0.1% Triton X-100 to permeabilize the cells. After incubation with 3% normal goat serum for 2 h at room temperature, fixed cells were incubated overnight at 4°C with antibodies against IBA1 (1:500, Cat#019-19741, WAKO, Japan) or p-Src-Tyr416 (1:200). Cells were washed with PBS and then incubated for 2 h with goat anti-rabbit IgG H&L (Alexa Fluor® 488) antibody (Cat#ab150077, 1:500, Abcam, United States) or goat anti-rabbit IgG H&L (Alexa Fluor® 647) antibody (Cat#ab150079, 1:500, Abcam, United States) followed by incubation with DAPI for 10 min. Coverslips were imaged by confocal microscopy (Leica, Germany). Quantification was carried out using ImageJ software.
5
Immunostaining of Ovarian Cancer Cells
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Εpithelial ovarian cancer cells were seeded on glass coverslips, and when they reached the 80% confluence, they were fixed with 100% methanol chilled at −20 °C for 5 min. Cells were treated with blocking solution (1% BSA, 22.52 mg·mL−1 glycine in PBS+ 0.1% Tween 20) to eliminate unspecific binding for 30 min at room temperature. Cells were incubated with primary anti‐MUC16 antibody (EPSISR23) diluted 1 : 150 overnight at 4 °C and then with the secondary Goat Anti‐Rabbit IgG H&L Alexa Fluor® 488 antibody (ab150077, ABCAM plc Cambridge, UK) diluted 1 : 1000 in 1% BSA for 1 h at RT protecting them from light. The glass coverslips were mounted using ProLong Diamond Antifade Mountant with DAPI, and the images were acquired using Cytation 5 cell imaging multimode reader with objective lens 20× and processed by gen.5 software (BioTek).
6
Primary Human Mesenchymal Stem Cell Culture
7
Immunofluorescence Staining of IgG
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Deparaffinization and rehydration were the same as those used for H&E staining. HIAR was performed using antigen retrieval buffer (Abcam, Cambridge, UK) at 98°C for approximately 20 minutes. The sections were incubated with Protein Block (Abcam, Cambridge, UK) for 1 hour at room temperature for tissue blocking. The sections were incubated overnight at 4°C with a rabbit anti-human IgG monoclonal antibody (Abcam, Cambridge, UK) at a ratio of 1:500, and then the sections were washed 3 times with PBS, followed by incubation with a goat anti-rabbit IgG H&L antibody (Alexa Fluor® 488) (Abcam, Cambridge, UK) at a ratio of 1:300 for 1 hour at room temperature. After washing, the sections were sealed and observed under an Olympus fluorescence microscope.
8
Comprehensive Immunofluorescence Staining of Embryos
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The protocol used for the immunofluorescence staining of embryos was previously described by our research group (Huang et al., 2019 (link); Li et al., 2019 (link)). The sections were visualized using an LSM 800 laser scanning microscope (ZEISS, Germany). F-actin was labeled with Alexa Fluor™ 488 phalloidin, which was purchased from Invitrogen (Carlsbad, CA, USA). A rabbit anti-gamma H2A.X antibody (phospho-S139), a rabbit anti-Aurora B antibody, DAPI staining solution and a goat anti-rabbit IgG H&L antibody (Alexa Fluor® 488) were obtained from Abcam (Cambridge, UK). An Alexa Fluor® 594-conjugated mouse anti-MAD2 antibody was obtained from Santa Cruz (Dallas, TX, USA). A mouse anti-α-tubulin-FITC monoclonal antibody was procured from Sigma-Aldrich (St. Louis, MO, USA).
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