Cells were plated on poly-d -lysine coated slides (Sigma Aldrich Oakville, ON, Canada), then fixed in 4% paraformaldehyde and permeabilized with 1 x PBS–0.2% Triton X-100. After blocking, cells were incubated with different primary antibodies, including anti-β-catenin (Santa-Cruz Biotechnology Dallas, TX, USA), anti-LY75 (Santa-Cruz Biotechnology Dallas, TX, USA and Abcam, Branford, CT, USA), anti-APC2 (Abcam, Branford, CT, USA), anti-Axin1 (Santa-Cruz Biotechnology Dallas, TX, USA), and anti-DSP (Santa-Cruz Biotechnology Dallas, TX, USA), and subsequently incubated with secondary antibodies, including rhodamine-linked goat-anti-mouse IgG1 (Santa Cruz Biotechnology Dallas, TX, USA) or Alexa Fluor 488-labeled goat anti-rabbit antibody (Abcam, Branford, CT, USA). Cells were finally stained with 4′,6-diamidino-2-phenylindole (DAPI). Images were captured using a Zeiss LSM 700 confocal microscope (Carl Zeiss Meditec AG Jena, Germany).
Alexa fluor 488 labeled goat anti rabbit antibody
Manufactured by Abcam
Sourced in United States
Alexa Fluor 488-labeled goat anti-rabbit antibody is a secondary antibody conjugated with the fluorescent dye Alexa Fluor 488. It is designed to bind to rabbit primary antibodies, allowing for detection and visualization of target proteins or biomolecules in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti ly75, by Santa Cruz Biotechnology (2 mentions)
Anti apc2, by Abcam (2 mentions)
Anti axin1, by Santa Cruz Biotechnology (2 mentions)
Rhodamine linked goat anti mouse igg1, by Santa Cruz Biotechnology (2 mentions)
Anti β catenin, by Santa Cruz Biotechnology (2 mentions)
Anti dsp, by Santa Cruz Biotechnology (1 mentions)
Poly d lysine coated slides, by Merck Group (1 mentions)
Lsm 700 confocal microscope, by Zeiss (1 mentions)
Anti vimentin, by Santa Cruz Biotechnology (1 mentions)
Anti e cadherin, by Santa Cruz Biotechnology (1 mentions)
Anti n cadherin, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti mouse cd206 antibody, by Abcam (1 mentions)
Rat anti mouse cd31 monoclonal antibody, by Abcam (1 mentions)
4 protocols using alexa fluor 488 labeled goat anti rabbit antibody
1
Immunostaining of Cell Signaling Proteins
2
Immunofluorescence Analysis of Cell Markers
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IF analysis was performed as previously described [26 (link)]. Fixed samples were incubated with different primary antibodies, including anti-β-catenin (Santa-Cruz Biotechnology Dallas, TX, USA), anti-LY75 (Santa-Cruz Biotechnology Dallas, TX, USA and Abcam, Branford, CT, USA), anti-APC2 (Abcam, Branford, CT, USA), anti-Axin1 (Santa-Cruz Biotechnology Dallas, TX, USA), anti-N-cadherin, anti-E-cadherin, anti-vimentin, anti-EpCAM, and subsequently incubated with secondary antibodies, including rhodamine-linked goat-anti-mouse IgG1 (Santa Cruz Biotechnology Dallas, TX, USA) or Alexa Fluor 488-labeled goat anti-rabbit antibody (Abcam, Branford, CT, USA).
3
Immunostaining Protocol for CCR7 and CD206
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The RAW264.7 cells, which had undergone different treatments, were washed three times with PBS for 3 min each time. They were then fixed with 4% paraformaldehyde for 15 min, followed by three additional PBS washes, each for 3 min. Permeabilization was achieved by incubating the cells with 0.5% Triton X‐100 at room temperature for 20 min, followed by another PBS wash. The blocking step was carried out by adding 5% BSA at room temperature for 30 min. After removing the blocking solution, an appropriate amount of either rabbit anti‐mouse CCR7 antibody (1:100) or rabbit anti‐mouse CD206 antibody (1:50, Abcam) was added, and the cells were incubated at 4 °C overnight. The following day, the primary antibodies were removed, and Alexa Fluor 488‐labeled goat anti‐rabbit antibody (1:500, Abcam) was added to bind to CCR7, along with Alexa Fluor 594‐labeled goat anti‐rabbit antibody (1:500, Abcam) to bind to CD206. After a 1‐h incubation, the cells were washed three times with PBS, and DAPI was added for light‐protected incubation for 5 min. CLSM was used for image acquisition.
4
Nanobubble-Mediated Tumor Targeting Evaluation
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Tumor-bearing nude mice were sacrificed at 8 minutes after injection of fluorescence-labeled nanobubbles (5×108/mL), and xenograft tumor tissues were collected. After injection of DiI-labeled targeted nanobubbles, the sections of xenograft tumor tissues were incubated with rat anti-mouse CD31 monoclonal antibody (1:300; Abcam, Cambridge, UK) at 4°C overnight and Alexa Fluor® 488-labeled rabbit anti-rat antibody (1:300; Abcam) for 1 hour. After being counterstained with DAPI, the sections were observed under CLSM to evaluate the distribution of targeted nanobubbles. To confirm the ability of targeted nanobubbles to load CAIX aptamer in vivo, the sections of xenograft tumor tissues after injection of 6-FAM and DiI-labeled targeted nanobubbles were stained with DAPI and observed under CLSM. The cryosections of xenograft tumor tissues after injection of DiI-labeled targeted or non-targeted nanobubbles were used to evaluate the binding ability of nanobubbles in xenograft tumor tissues. The sections were incubated with rabbit anti-human CAIX polyclonal antibody (1:200; Abcam) at 4°C overnight, then incubated with Alexa Fluor® 488-labeled goat anti-rabbit antibody (1:300; Abcam) for 1 hour, and observed under CLSM.
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