Alexa fluor 488 goat anti rabbit igg

Manufactured by Abcam

Sourced in United States, United Kingdom, Germany

Alexa Fluor 488 goat anti-rabbit IgG is a secondary antibody conjugated with Alexa Fluor 488 dye. It is designed to detect and visualize rabbit primary antibodies in various immunoassays and imaging applications.

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Lab products found in correlation

Alexa fluor 594 goat anti mouse igg, by Abcam (8 mentions) 4 6 diamidino 2 phenylindole (dapi), by Beyotime (7 mentions) Ab150077, by Abcam (6 mentions) Triton x 100, by Merck Group (5 mentions) Alexa fluor 555 goat anti mouse igg, by Abcam (4 mentions) Bovine serum albumin (bsa), by Merck Group (4 mentions) Triton x 100, by Solarbio (4 mentions) Lsm 510 meta, by Zeiss (4 mentions) Paraformaldehyde (pfa), by Solarbio (4 mentions) Alexa fluor 647 goat anti rat igg, by Abcam (3 mentions) Alexa fluor 488 goat anti rat igg, by Abcam (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Solarbio (3 mentions) Alexa fluor 555 goat anti rabbit igg, by Abcam (3 mentions) F4 80, by Abcam (3 mentions) Lsm 880, by Zeiss (3 mentions) Alexa fluor 647 goat anti mouse igg, by Abcam (2 mentions) Alexa fluor 594 goat anti rat igg, by Abcam (2 mentions) Laser scanning confocal microscopy, by Leica Biosystems (2 mentions) Mab377, by Merck Group (2 mentions) Alexa fluor 568 goat anti mouse igg, by Abcam (2 mentions)

91 protocols using alexa fluor 488 goat anti rabbit igg

Adipose Tissue Immunofluorescence Staining

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Tissue sections were deparaffinized, rehydrated, and washed three times with Tris-buffered saline (Sigma). Sections were incubated with H₂O₂ for 30 min and washed with distilled water to avoid nonspecific background staining. Adipose tissue samples were stained with anti-CD34 (Abcam) and guinea pig anti‐mouse perilipin (Progen) antibodies following the manufacturers’ instructions. After washing, the samples were incubated with goat anti-rabbit Alexa Fluor® 488 IgG (Abcam) and goat anti-guinea pig Alexa Fluor® 647 IgG (Abcam). Nuclei were stained with DAPI (Sigma, USA).
For migrasomes detection, tissue samples were stained with anti-TSPAN4 (Biossusa), anti-TSPAN7 (Biorbyt) antibodies following the manufacturers’ instructions and then with goat anti-rabbit Alexa Fluor® 488 IgG (Abcam) and goat anti-rabbit Alexa Fluor® 647 IgG (Abcam). Nuclei were stained with DAPI (Sigma, USA). Furthermore, the tissue samples were stained with anti-integrin β1 (SantaCruz), and then with goat anti-mouse Alexa Fluor® 488 IgG (Abcam). Nuclei were stained with DAPI (Sigma). Images were obtained using Axio Image D2 microscope (Zeiss) and analyzed using ImageJ software.

Chen Y., Li Y., Li B., Hu D., Dong Z, & Lu F. (2024). Migrasomes from adipose derived stem cells enrich CXCL12 to recruit stem cells via CXCR4/RhoA for a positive feedback loop mediating soft tissue regeneration. Journal of Nanobiotechnology, 22, 219.

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Phospho-specific GPR35 Antibody Characterization

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The rabbit phospho-site–specific GPR35 antiserum pSer³⁰⁰/pSer³⁰³-hGPR35a (Cat number (7TM0102C), raised against the sequence KAHKpSQDpSLCVTL, and the pSer²⁹⁸/pSer³⁰¹-mGPR35 antiserum (7TM0102B), raised against the sequence TPHKpSQDpSQILSLT, were developed in collaboration with 7TM Antibodies GmbH. IRDye 800CW donkey anti-rabbit IgG, IRDye 800CW donkey anti-goat IgG, and IRDye 800CW goat anti-rat IgG were from LI-COR Biosciences. Alexa Fluor 488-goat anti-rabbit IgG and Alexa Fluor 594-donkey anti-rat IgG were from Abcam. Mouse monoclonal anti-FLAG M2 was from Merck. Horseradish peroxidase anti-mouse (sheep) was from GE Healthcare. High affinity anti-HA (rat) and anti-HA affinity matrix were from Roche Diagnostics.

Divorty N., Jenkins L., Ganguly A., Butcher A.J., Hudson B.D., Schulz S., Tobin A.B., Nicklin S.A, & Milligan G. (2022). Agonist-induced phosphorylation of orthologues of the orphan receptor GPR35 functions as an activation sensor. The Journal of Biological Chemistry, 298(3), 101655.

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Immunocytochemical Profiling of hWJMSCs

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Cells were fixed with 4% paraformaldehyde, blocked with normal goat serum and incubated with primary antibodies. The primary antibodies used were a mouse monoclonal antibody against S100β (1:1000, Abcam), a rabbit polyclonal antibody against low affinity nerve growth factor receptor (p75 NGFR) (1:1000, Abcam), a mouse monoclonal antibody against myelin binding protein (MBP) (1:1000, Thermo Fisher), a mouse monoclonal antibody against glial fibrillary acidic protein (GFAP) (2 μg/ml, Stem Cell™ Technologies) and a mouse monoclonal antibody against myelin oligodendrocyte glycoprotein (MOG) (2 μg/ml, Abcam). Alexa Fluor® 488 goat anti-rabbit IgG (1:300, Abcam) and Texas Red® goat anti-mouse IgG (1:300, Abcam) were used as the secondary antibodies. An internal control for the assay was included by omitting the primary antibodies from the samples. Cell nuclei were counterstained with 4′, 6-diamidino-2-phenylindole (DAPI). The cells in various groups were examined by fluorescence microscopy (Nikon A1_R, Japan) using the same laser intensity and detection sensitivity. The expression of all tested neural markers in differentiated and undifferentiated hWJMSCs was analyzed qualitatively and compared to the positive control.

Omar N., Lokanathan Y., Mohd Razi Z.R, & Bt Haji Idrus R. (2019). The effects of Centella asiatica (L.) Urban on neural differentiation of human mesenchymal stem cells in vitro. BMC Complementary and Alternative Medicine, 19, 167.

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Immunofluorescence Staining of A-375 Cells

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A-375 cells were fixed with 4% paraformaldehyde for 20 min at 37 °C and permeabilized with 0.1% Triton X-100 for 5 min at room temperature. Cells were blocked with 3% bovine serum albumin (BSA) for 1 h and then incubated overnight with the primary antibody. Next day, coverslips were washed and probed for 1 h with Alexa Fluor 488 goat anti-rabbit IgG (Abcam, Cambridge, United Kingdom). After washing, the cells were incubated with DAPI for 5 min. Coverslips were mounted with Vectashield and captured using an Olympus BX41 microscope, images were analyzed with CellSens software (Olympus, Shinjuku, Japan).

Krejcir R., Krcova L., Zatloukalova P., Briza T., Coates P.J., Sterba M., Muller P., Kralova J., Martasek P., Kral V, & Vojtesek B. (2019). A Cyclic Pentamethinium Salt Induces Cancer Cell Cytotoxicity through Mitochondrial Disintegration and Metabolic Collapse. International Journal of Molecular Sciences, 20(17), 4208.

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CXCR4 Receptor Immunofluorescence Staining

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The CXCR4 receptor was identified on cells by immunofluorescence staining. Cells were plated in a fibronectin coated 24 well plate at a density of 5000 cells/cm² and incubated in 500 μl culture medium for 24 hours at 37°C. Cells were fixed in 4% paraformaldehyde for 40 minutes, blocked with 1% BSA overnight at 4°C, incubated with rabbit anti-CXCR4 (Thermo Scientific) (1:200) primary antibody for 45 minutes at 37°C, and incubated with AlexaFluor 488 goat anti-rabbit IgG (Abcam) (1:400) secondary antibody for 45 minutes at 37°C. Nuclei were counterstained with DAPI.

Warner H., Wu Y, & Wagner W.D. (2021). Syndecan-4 functionalization of tissue regeneration scaffolds improves interaction with endothelial progenitor cells. Regenerative Biomaterials, 8(6), rbab070.

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Angiogenesis and Inflammation Analysis

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After being embedded in paraffin, samples that were harvested at 2 and 12 weeks were stained with Von Willebrand factor (vWF) antibody (1:250, Abcam), and some of them that were harvested after 5 days of implantation were stained with CD68 (1:500, Abcam) and anti‐perilipin antibody (1:250, Abcam), followed by staining with Alexa Fluor 488 goat anti‐rabbit IgG (1:500, Abcam) as the secondary antibody. Cell nuclei were stained using 4′,6‐diamidino‐2‐phenylindole. Next, vWF‐positive regions, which indicated the vessels; perilipin‐positive areas, which indicated intact adipocytes; and CD68‐positive macrophages were counted under a high‐magnification field (average from 10 high‐magnification fields per section).

Lv T., Gu Y., Bi J., Kang N., Yang Z., Fu X., Wang Q., Yan L., Liu X., Cao Y, & Xiao R. (2019). Fructose 1,6‐Bisphosphate as a Protective Agent for Experimental Fat Grafting. Stem Cells Translational Medicine, 8(6), 606-616.

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Immunofluorescence Staining of Macrophages

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Raw264.7 cells and BMDMs were fixed with 4% paraformaldehyde for 15 min, followed by permeabilization for 10 min. After blocking with 5% BSA, the cells were incubated with primary antibodies against iNOS (1 : 200, ab178945, Abcam), Arg1 (1 : 50, #93668S, Cell Signaling Technology), and F4/80 (1 : 50, MAB5580-SP, R&D systems, USA) overnight at 4°C. The next day, corresponding secondary antibodies (1 : 500, Alexa Fluor® 488 goat anti-rabbit IgG and Alexa Fluor® 594 goat anti-mouse IgG, Abcam) were applied, followed by the nuclei staining with 4′,6-diamidino-2-phenylindole (DAPI). Confocal lazer scanning analysis of Raw264.7 cells and BMDMs was performed using a laser confocal microscope (Zeiss LSM880, Carl Zeiss AG, Germany).

Xu M.M., Kang J.Y., Ji S., Wei Y.Y., Wei S.L., Ye J.J., Wang Y.G., Shen J.L., Wu H.M, & Fei G.H. (2022). Melatonin Suppresses Macrophage M1 Polarization and ROS-Mediated Pyroptosis via Activating ApoE/LDLR Pathway in Influenza A-Induced Acute Lung Injury. Oxidative Medicine and Cellular Longevity, 2022, 2520348.

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Immunofluorescence Characterization of Stem Cells and Cardiomyocytes

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Cells were seeded on a coverslip and cultured for 3 to 5 days before being fixed in 4% paraformaldehyde for 20 minutes. The cells were permeabilized with 0.1% Triton X-100 for 5 minutes when needed. Cells were then blocked in 5% BSA in PBS for 30 minutes; all these procedures were performed at room temperature. Cells were then incubated overnight at 4 °C with primary antibodies. For pluripotent stem cells, anti-SSEA4 (1:200 dilution; Abcam, UK) and anti-Nanog (1:200 dilution; Abcam) were applied. For cardiomyocytes, anti-α-actinin (1:200 dilution; Sigma-Aldrich), anti-cardiac troponin I (1:200 dilution; Abcam), and anti-Nav1.5 (1:100 dilution; Abcam) were used. The following secondary antibodies included Alexa Fluor 488 goat anti-rabbit IgG (1:800 dilution; Abcam) and Alexa Fluor 555 goat anti-mouse IgG (1:800 dilution; Abcam) were incubated with samples at room temperature for 30 minutes. The nuclei were stained with DAPI (Life Technologies, USA). Then, the cells were rinsed with PBS and observed under a fluorescence microscope (Axio Imager A2, Carl Zeiss, Germany). Images were analyzed and merged with ImageJ (NIH, version 1.8.0_77).

Zhu Y., Wang L., Cui C., Qin H., Chen H., Chen S., Lin Y., Cheng H., Jiang X, & Chen M. (2021). Pathogenesis and drug response of iPSC-derived cardiomyocytes from two Brugada syndrome patients with different Nav1.5-subunit mutations. Journal of Biomedical Research, 35(5), 395-407.

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Exosome-Mediated Regulation of HSC Phenotype

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P6 HSC were treated for 36 hrs with or without exosomes (8 μg/ml) from D1-3 HSC in the presence or absence of anti-integrin αvβ3 or anti-integrin α5β1. Cells were fixed and incubated with NH1 anti-CCN2 IgY (5μg/ml [25 (link)]), anti-αSMA (1:100, Dako Cytomatio, Denmark) or anti-collagen α(1) (1:250, Abcam), followed by Alexa Fluor® 568 goat-anti chicken IgY, Alexa Fluor® 647 goat-anti mouse IgG, or Alexa Fluor® 488 goat-anti rabbit IgG, respectively. The cells were mounted with Vectashield Mounting Medium containing 4',6-diamidino-2-phenylindole (DAPI) nuclear stain (Vector Laboratories, Burlingame, CA), and examined by confocal microscopy.

Chen L, & Brigstock D.R. (2016). Integrins and heparan sulfate proteoglycans on hepatic stellate cells (HSC) are novel receptors for HSC-derived exosomes. FEBS letters, 590(23), 4263-4274.

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Exosome Uptake and HSP60 Localization

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The labeled exosomes (10 μg/mL) were incubated with MH-S cells. After 12 h, the cells were fixed with 4% paraformaldehyde, washed with PBS, blocked in 3% BSA for 1 h and reacted with primary antibody against rabbit anti-HSP60 (1:200, Proteintech, USA) overnight. The cover glass was further stained with Alexa Fluor 488 goat anti-rabbit IgG (1:1000, Abcam, USA) for 1 h in the dark. After being washed with PBS, the slices were stained with DAPI (1:1000, Beyotime, Shanghai, China) for 10 min, washed with PBS, and added the anti-fluorescence quencher before mounting onto microscope slides. Finally, cells were observed using Leica TCS SP8 laser scanning confocal microscope (Leica Microsystems Ltd., Wetzlar, Germany). Images were analyzed using LAS X software.

Xia L., Zhang C., Lv N., Liang Z., Ma T., Cheng H., Xia Y, & Shi L. (2022). AdMSC-derived exosomes alleviate acute lung injury via transferring mitochondrial component to improve homeostasis of alveolar macrophages. Theranostics, 12(6), 2928-2947.

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