Alexa fluor 488 labeled donkey anti rabbit igg

Manufactured by Thermo Fisher Scientific

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Alexa Fluor 488-labeled donkey anti-rabbit IgG is a secondary antibody conjugated with the Alexa Fluor 488 fluorescent dye. It is designed to detect and visualize rabbit primary antibodies in various immunoassay applications.

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Lab products found in correlation

Alexa fluor 594 labeled donkey anti mouse igg, by Thermo Fisher Scientific (4 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (3 mentions) Dapi staining solution, by Solarbio (2 mentions) Lsm 710, by Zeiss (2 mentions) Mouse anti β actin, by Cell Signaling Technology (1 mentions) Rabbit anti p p38 mapk, by Cell Signaling Technology (1 mentions) Incomplete freund s adjuvant, by Merck Group (1 mentions) Complete freund s adjuvant, by Merck Group (1 mentions) Ab5096, by Abcam (1 mentions) Ab13840, by Abcam (1 mentions) Normal donkey serum (nds), by Merck Group (1 mentions) Hoechst 33342, by Dojindo Laboratories (1 mentions) Bz x710 microscope, by Keyence (1 mentions) Sabpo kit, by Nichirei Biosciences (1 mentions) Bz x710 all in one fluorescence microscope, by Keyence (1 mentions) Mounting medium, by Vector Laboratories (1 mentions) Rabbit anti wt1, by Santa Cruz Biotechnology (1 mentions) 510 confocal microscope, by Zeiss (1 mentions) Rabbit anti gfp antibody, by Thermo Fisher Scientific (1 mentions) Alexa fluor 546 labeled donkey antigoat igg, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Alexa Fluor 488 (6806 citations) Alexa 488 (1863 citations) Alexa Fluor 488 anti-rabbit (273 citations) Alexa Fluor 488 donkey anti-rabbit IgG (272 citations) Alexa Fluor 488 donkey anti-rabbit (245 citations) Donkey anti-rabbit Alexa 488 (85 citations) Donkey anti-rabbit 488 (44 citations) Alexa Fluors (21 citations) Donkey anti-rabbit IgG Alexa 488 (18 citations) Alexa Fluor 488-labeled anti-rabbit (9 citations)

17 protocols using alexa fluor 488 labeled donkey anti rabbit igg

Antibody Generation for CUB-Serine Protease

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For CUB-serine protease (CUB-SP), one segment from the CUB domain (amino acids 78–178) and one from the serine protease domain (amino acids 398–498) were selected for antibody generation. These two segments were fused into the pET-28a and pGEX-4T vectors, respectively, for over-expression. His₆-tagged protein was injected 4 times intracutaneously into the backs of New Zealand white rabbits at 3-week intervals. The first two injections were mixed with Freund’s complete adjuvant (Sigma, USA), and the last two injections were mixed with Freund’s incomplete adjuvant (Sigma, USA). Sera were collected from the rabbits at 1 week after the final injection, and antibody was purified using affinity purification with GST-tagged protein as the ligands.
Rabbit anti-pp38 MAPK (phospho-p38MAPK, Thr180/Tyr182) and mouse anti-β-actin, as well as HRP-linked goat anti-rabbit and anti-mouse IgG secondary antibodies were purchased from Cell Signaling Technology (USA). Mouse anti-PKAα/β and rabbit anti-serotonin was purchased from Santa Cruz Biotechnology, Inc. (USA) and Sigma Aldrich (USA), respectively. Alexa Fluor 488-labeled donkey anti-rabbit IgG and Alexa Fluor 594-labeled donkey anti-mouse IgG secondary antibodies were purchased from Life Technologies (USA).

Zhang G., He L.S., Him Wong Y., Xu Y., Zhang Y, & Qian P.Y. (2015). p38 MAPK regulates PKAα and CUB-serine protease in Amphibalanus amphitrite cyprids. Scientific Reports, 5, 14767.

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Oocyte and FEC Marker Analysis in Rodent CRs

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The oocyte marker DEAD (Asp-Glu-Ala-Asp) box polypeptide 4 (DDX4) and the FEC marker fork head box L2 (FOXL2) were analyzed to determine whether the oocytes of CRs, in particular those in MOFs or DNOs, showed biological characteristics similar to those of other rodents. Briefly, DDX4 and FOXL2 were expressed in germ cells and FECs from the time of their first appearance to adulthood in mice (Yamashita et al., 2015 (link)). Deparaffinized sections were treated with 10 mM citrate buffer for 20 min at 105°C for antigen retrieval. These sections were then blocked with 5% normal donkey serum (Sigma-Aldrich, St. Louis, MO, United States). They were incubated overnight with rabbit anti-DDX4 (1:500 dilution; ab13840; Abcam, Cambridge, United Kingdom) and goat anti-rabbit FOXL2 antibody (1:1000; ab5096; Abcam) at 4°C. The sections were incubated for 1 h at room temperature with Alexa Fluor-488 labeled donkey anti-rabbit IgG (1:500, Life Technologies, Carlsbad, CA, United States) and Alexa Fluor-564 labeled donkey anti-goat IgG (1:500, Life Technologies) for DDX4 and FOXL2 double immunofluorescence. The sections were counterstained with Hoechst 33342 (1:200; Dojindo, Kumamoto, Japan). The stained sections were examined using a BZ-X710 microscope (Keyence).

Islam M.R., Ichii O., Nakamura T., Irie T., Masum M.A., Otani Y., Namba T., Chuluunbaatar T., Elewa Y.H, & Kon Y. (2021). Developmental Changes of the Ovary in Neonatal Cotton Rat (Sigmodon hispidus). Frontiers in Physiology, 11, 601927.

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Histopathological and Immunodetection Analysis in Kidney

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The tissue sections were stained with periodic acid Schiff-hematoxylin (PAS-H) for histopathological analysis. Immunodetection of target cell markers was performed using kidney specimens fixed with PFA; details are listed in Supplementary Table 1.
Briefly, deparaffinization and antigen retrieval was followed by submerging tissue sections in methanol containing 3% H2O2 for 20 min at room temperature and blocked with normal goat or donkey serum. Sections were incubated with primary antibody overnight at 4°C followed by incubation with the respective secondary antibodies at room temperature for 30 min: Alexa Fluor 546-labeled donkey anti-mouse IgG (Life Technologies, Yokohama, Japan) or Alexa Fluor 488-labeled donkey anti-rabbit IgG (Life Technologies, Yokohama, Japan). The sections were examined under an All-in-One Fluorescence Microscope BZ-X710 (Keyence, Osaka, Japan).
For immunohistochemistry, the sections were incubated with the appropriate biotinylated secondary antibody for 30 min, then with streptavidin-horseradish peroxidase (SABPO kit; Nichirei, Tokyo, Japan) for another 30 min, followed by incubation with 3,3-diaminobenzidine tetrahydrochloride-H2O2 solution. Finally, the sections were counterstained with hematoxylin, dehydrated in an ascending series of alcohol solutions, and cleared with xylene.

Masum M.A., Ichii O., Ichii O., ELewa Y.H.A, & Kon Y. (2019). Induced expression of Toll-like receptor 9 in peritubular capillary endothelium correlates with the progression of tubulointerstitial lesions in autoimmune disease-prone mice. Lupus, 28(3).

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Imaging GFP Expression in Kidney Tissue

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Ten micrometer frozen sections of formalin-fixed tissues were treated with 0.1% Triton X-100. Blocking was performed with 5% goat serum plus 5% bovine serum albumin followed by overnight incubation with rabbit anti-GFP antibody (1:400; Invitrogen). Anti-GFP antibodies were detected by fluorescence-conjugated goat antirabbit antibody IgG Alexa Fluor 488 (1:500; Invitrogen) and treatment with mounting medium (Vector). Tissues were then examined with a Zeiss 510 confocal microscope using FITC fluorescence filter (Carl Zeiss Microimaging, Thornwood, NY).
To confirm the location of the GFP expression in the kidney, multiple immunofluorescence study was performed using a podocyte marker, rabbit anti-wt1 (1:200; Santa Cruz), and goat anti-GFP (1:400; Invitrogen) antibody overnight at 4 °C, followed by incubation with Alexa Fluor 488-labeled donkey antirabbit IgG (1:200; Invitrogen) and Alexa Fluor 546-labeled donkey antigoat IgG (1:200; Invitrogen) at room temperature for 2 hours. Sections were washed, mounted with VECTASHIELD mounting medium with DAPI (4’,6-diamidino-2-phenylindole) (Vector Laboratories), and then examined by laser confocal microscopy (LSM 710; Carl Zeiss AG).

Picconi J.L., Muff-Luett M.A., Wu D., Bunchman E., Schaefer F, & Brophy P.D. (2014). Kidney-specific expression of GFP by in-utero delivery of pseudotyped adeno-associated virus 9. Molecular Therapy. Methods & Clinical Development, 1, 14014-.

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Immunohistochemical and Immunofluorescent Tissue Analysis

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Immunohistochemistry and immunofluorescence were performed using formalin-fixed, paraffin-embedded tissues. Four-μm-thick sections were cut and dewaxed in xylene, rinsed in graded ethanol, and rehydrated in distilled water. After antigen retrieval with EDTA buffer (1 mM Tris/EDTA, pH 9.0), endogenous peroxidase activity was blocked with 3% H2O2. Then slides were incubated with primary antibody (PDPN, 1:100, Abcam, ab236529, USA; CD163, Abcam, ab156769, USA; CD18, Affinity, BF0227, China). For IHC, markers were detected with a Goat Anti-rabbit IgG Two-step Detection Kit (PV-9000, ZSGB-Bio, China). Next, the slides were counterstained with Mayer Hematoxylin Solution (G1080, Solarbio, China) for nuclear staining. For IF, Alexa-Fluor 488 labeled donkey anti-rabbit IgG (Invitrogen, USA, 1:1000) and Alexa-Fluor 594 labeled donkey anti-mouse IgG (Invitrogen, USA, 1:1000) were applied to the double-colored fluorescent staining. Nucleus was labeled by DAPI staining solution (Solarbio, China).

Wang X., Wang X., Li J., Liang J., Ren X., Yun D., Liu J., Fan J., Zhang Y., Zhang J., Ren X., Zhang H., Shang G., Sun J., Chen L., Chen L., Li T., Tong L., Zhang C., Yu S, & Yang X. (2022). PDPN contributes to constructing immunosuppressive microenvironment in IDH wildtype glioma. Cancer Gene Therapy, 30(2), 345-357.

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Immunofluorescence Staining of Cardiac Fibroblasts

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The CFs were fixed with 4% paraformaldehyde at room temperature for 20 min. Then they were permeabilized with 0.1% Triton X-100 for 15 min and blocked with 5% BSA for 30 min. The CFs were then incubated with the primary antibody α-SMA (1:200, 19245, CST) and Ki67 (1:200, A23722; ABclonal, Woburn, MA, USA) at 4 °C overnight. The next day, after washing with PBS, the CFs were incubated with Alexa Fluor^® 488 Labeled Donkey Anti-rabbit IgG (A-21206; Invitrogen, Waltham, MA, USA) for two hours at room temperature, in darkness. The nuclei were stained with 4-6-diamidino-2-phenylindole (DAPI; Invitrogen). The imaging was conducted using a laser confocal microscope (Nikon, Tokyo, Japan). The results were quantified by Image-Pro Plus 6.0 software (Media Cybernetics).

Zhang Q., Zhang Z., Chen W., Zheng H., Si D, & Zhang W. (2023). Rivaroxaban, a direct inhibitor of coagulation factor Xa, attenuates adverse cardiac remodeling in rats by regulating the PAR-2 and TGF-β1 signaling pathways. PeerJ, 11, e16097.

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Retinal Explant Immunostaining Protocol

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Rat retinal explants were fixed with 4% PFA and plus 4% sucrose and snap frozen in OCT (VWR Chemicals, UK). Cryosections were prepared, washed in PBS and permeabilized with 0.1% Triton X-100. Non-specific staining was blocked with 5% BSA in PBS for 30 min at RT followed by immunostaining with goat anti-mouse GFAP (Cell Signaling Technology, UK, 1:100), rabbit anti-mouse Brn-3a (Santa Cruz Biotechnology, Inc., UK, 1:50), goat anti-mouse Rhodopsin (Abcam, UK, 1:100) antibodies. The secondary antibodies Alexa Fluor 549-labeled donkey anti-goat IgG (Invitrogen, CA) and Alexa Fluor 488-labeled donkey anti-rabbit IgG (Invitrogen, CA), both with 1:200 dilution in 2% BSA in PBS at room temperature (RT) for 1 h in the dark. Exclusion of the primary antibody acted as the negative control.

Ou K., Mertsch S., Theodoropoulou S., Wu J., Liu J., Copland D.A., Schrader S., Liu L, & Dick A.D. (2019). Restoring retinal neurovascular health via substance P. Experimental Cell Research, 380(2), 115-123.

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Immunohistochemical Analysis of Lung Tissue

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H&E and Alcian blue (AB) stains were performed on lung sections as described [35] (link). For IHC, monoclonal antibody against eosinophil specific major basic protein (anti-MBP, a kind gift from Drs. Nancy and James J. Lee, Mayo Clinic, Scottsdale, AZ) was used (1∶500) for eosinophils and anti-IL-22 antibody was used to detect IL-22 (1∶180). ABC staining kits were used to amplify the signal (Santa Cruz Biotechnology). Immunofluorescence of p-STAT3 was performed with rabbit anti-mouse phospho-Stat3 (Tyr705) (Cell Signaling, Danvers, MA) and Alexa Fluor 488-labeled donkey anti-rabbit IgG as secondary antibody (Invitrogen) and DAPI for nuclei. Tissue sections were mounted and examined using Zeiss LSM 510 laser scanning confocal microscope (Carl Zeiss) at 350 nm to assess p-STAT3 and 405 nm to assess cell nuclei.

Fang P., Zhou L., Zhou Y., Kolls J.K., Zheng T, & Zhu Z. (2014). Immune Modulatory Effects of IL-22 on Allergen-Induced Pulmonary Inflammation. PLoS ONE, 9(9), e107454.

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Immunofluorescence Staining of Tet1 and γH2AX

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Cells were fixed with 4% paraformaldehyde for 20 min at room temperature, washed three times with PBS (5 min each time), permeabilized with 0.25% Triton X-100 for 10 min at room temperature, washed with PBS (three times, 5 min each), and blocked with 5% BSA for 60 min at 37 ℃. The cells were then incubated with the following primary antibodies: rabbit anti-Tet1 (1:500, Abcam) and rabbit anti-H2AX (1:400, Abcam), overnight at 4 ℃. Cells were washed with PBS and probed with Alexa Fluor 488-labeled donkey anti-rabbit IgG (1:1000; Invitrogen) for 60 min at 37 ℃. After three PBS washes (5 min each), the nuclei were stained with DAPI for 10 min. Five microscope fields (× 200) were selected randomly for the evaluation. The mean fluorescence intensity was analysed using the ImageJ software.

Yao D., Zhao J., Zhang Q., Wang T., Ni M., Qi S., Shen Q., Li W., Li B., Ding X, & Liu Z. (2022). Aberrant methylation of Serpine1 mediates lung injury in neonatal mice prenatally exposed to intrauterine inflammation. Cell & Bioscience, 12, 164.

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Cerebellum Development Immunohistochemistry

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After perfusion fixation with Bouin's solution, cerebelli from 0.5 to 21-day-old mice were immersed in the same solution overnight or for 1 day. Sagittal sections of 6 μm in thickness were used for morphological analyses. Hematoxylin-eosin (H&E) staining was performed with paraffin sections. Immunostaining with antibodies against c-Ret (1:150; Immuno Biological Laboratories), phosphorylated c-Ret Y1062 (1:50; Abcam), and Shh (1:100; Santa Cruz, H-160) diluted with Can Get Signal immunostaining solution (TOYOBO) was performed with paraffin and frozen sections (19 (link), 45 ). Immunostaining with antibodies against GABRA6 (1:1000; Chemicon), calbindin D28k (1:150; Chemicon), BLBP (1:200, Abcam), PAX6 (1:500, Abcam), and Ki67 (1:500, Abcam) was performed with paraffin sections. The VECTASTAIN Elite ABC kit (Vector), Envision kit/HRP (diaminobenzidine; DAB) (DAKO) with counterstaining of hematoxylin and Alexa Fluor 488-labeled donkey anti-rabbit IgG (1:1000, Invitrogen) with counterstaining of 4′, 6-diamidino-2-phenylindole (DAPI) were used. We validated the primary antibodies used in this study with no positive signals in the specimens processed under the same staining condition except for incubation without primary antibodies.

Ohgami N., Iizuka A., Hirai H., Yajima I., Iida M., Shimada A., Tsuzuki T., Jijiwa M., Asai N., Takahashi M, & Kato M. (2021). Loss-of-function mutation of c-Ret causes cerebellar hypoplasia in mice with Hirschsprung disease and Down's syndrome. The Journal of Biological Chemistry, 296, 100389.

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