Alexa fluor 594 conjugated goat anti rabbit igg h l

Manufactured by Thermo Fisher Scientific

Sourced in United States

Alexa Fluor 594-conjugated goat anti-rabbit IgG (H + L) is a secondary antibody used for the detection and visualization of rabbit primary antibodies. The antibody is conjugated with the Alexa Fluor 594 fluorescent dye, which emits red fluorescence when excited at the appropriate wavelength. This product is suitable for a variety of immunoassay techniques, including immunofluorescence, flow cytometry, and Western blotting.

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Lab products found in correlation

Alexa fluor 488 conjugated goat anti mouse igg h l, by Thermo Fisher Scientific (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Solarbio (2 mentions) Anti asc antibody, by Cell Signaling Technology (1 mentions) Rabbit anti mouse f4 80 antibody, by Abcam (1 mentions) Bovine serum albumin (bsa), by Beyotime (1 mentions) Paraformaldehyde (pfa), by Beyotime (1 mentions) Mouse anti flag mab, by Merck Group (1 mentions) Alexa fluor 488 conjugated goat anti rabbit igg h l, by Thermo Fisher Scientific (1 mentions) Rabbit anti flag pab, by Merck Group (1 mentions) Alexa fluor 568 conjugated goat anti mouse igg h l, by Thermo Fisher Scientific (1 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions) Ab180573, by Abcam (1 mentions) Mouse monoclonal anti gapdh, by Transgene (1 mentions) Fitc conjugated goat anti mouse igg, by Transgene (1 mentions) Anti flag m2 affinity gel, by Merck Group (1 mentions) S8639, by Selleck Chemicals (1 mentions) Hrp conjugated secondary antibody, by Affinity Biosciences (1 mentions) Il 1β, by Affinity Biosciences (1 mentions) Nlrp3, by Abcam (1 mentions) Gapdh, by Affinity Biosciences (1 mentions)

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10 protocols using alexa fluor 594 conjugated goat anti rabbit igg h l

Immunofluorescence Assay for Macrophage Detection

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PMs were transfected with CLP/siRNA as described above. After 48 hours, PMs were washed twice with cooled PBS and then fixed in paraformaldehyde (Beyotime Biotechnology, China) for 30 min at room temperature. Subsequently, PMs were blocked with 5% bovine serum albumin (BSA) (Beyotime Biotechnology, China) for 2 hours and incubated overnight at 4°C with anti-ASC antibody (Cell Signaling Technology, USA). The next day, cells were incubated with Alexa Fluor^TM 594-conjugated goat anti-rabbit IgG (H+L) (Invitrogen, ThermoFisher Scientific, USA) for 1 hour, followed by staining with DAPI (Solarbio LIFE SCIENCES, China) for 10 min. Finally, fluorescence intensity and density were detected by confocal fluorescence microscopy.
Macrophage content in frozen sections of colonic tissue was also detected by IF, as described in previous research.52 (link) The primary antibody was rabbit anti-mouse F4/80 antibody (Abcam, England), and the secondary antibody was Alexa Fluor^TM 594-conjugated goat anti-rabbit IgG (H+L) (Invitrogen, ThermoFisher Scientific, USA).

Huang J., Dai M., He M., Bu W., Cao L., Jing J., Cao R., Zhang H, & Men K. (2023). Treatment of Ulcerative Colitis by Cationic Liposome Delivered NLRP3 siRNA. International Journal of Nanomedicine, 18, 4647-4662.

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Culturing HEK293T cells and PAMs for ASFV research

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HEK293T cells and PAMs, which were stored in our laboratory, were cultured in RPMI 1640 medium (catalog no. C11875500BT; Gibco) supplemented with 5% antibiotics-antimycotics (10,000 IU/mL penicillin, 10,000 μg/mL streptomycin, and 25 μg/mL amphotericin B) (catalog no. 15240-062; Gibco) and 10% heat-inactivated fetal bovine serum (FBS) (catalog no. 10099-141C; Gibco) in a 37°C incubator in 5% CO₂. The ASFV pig/Heilongjiang/2018 (ASFV HLJ/2018) strain was isolated from field samples from China as described previously (63 (link)). ASFV-ΔH240R was generated in our previous study (8 (link)).
A rabbit anti-Myc PAb (catalog no. ab9106; Sigma-Aldrich), a mouse anti-Flag MAb (catalog no. ab62928; Sigma-Aldrich), and a rabbit anti-Flag PAb (catalog no. ab1162; Sigma-Aldrich) were purchased from Sigma-Aldrich. Alexa Fluor 488-conjugated goat anti-rabbit IgG (H+L) (catalog no. 2072687; Invitrogen), Alexa Fluor 488-conjugated goat anti-mouse IgG (H+L) (catalog no. 1942237; Invitrogen), Alexa Fluor 594-conjugated goat anti-rabbit IgG (H+L) (catalog no. 111-585-003; Invitrogen) and Alexa Fluor 568-conjugated goat anti-mouse IgG (H+L) (catalog no. 175697; Invitrogen) antibodies were purchased from Invitrogen. A mouse anti-GST MAb (catalog no. K200006M; Solarbio) and 4′,6-diamidino-2-phenylindole (DAPI) (catalog no. C006; Solarbio) were purchased from Solarbio.

Zhou P., Dai J., Zhang K., Wang T., Li L.F., Luo Y., Sun Y., Qiu H.J, & Li S. (2022). The H240R Protein of African Swine Fever Virus Inhibits Interleukin 1β Production by Inhibiting NEMO Expression and NLRP3 Oligomerization. Journal of Virology, 96(22), e00954-22.

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Purification and Characterization of Anti-Influenza Antibodies

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Hybridoma cells secreting mouse monoclonal anti-influenza A virus (IAV) NP (clone H16-L10-4R5, ATCC HB-65) and anti-IAV M (M2-1C6-4R3, ATCC HB-64) were obtained from ATCC and antibodies in the supernatant purified using a protein G column (Yewdell et al., 1981 (link)). Other antibodies used include: rabbit monoclonal anti-eIF4A3 (ab180573, Abcam), mouse monoclonal anti-IAV NP (M100014, Zoonogen, China), rabbit polyclonal anti-IAV M1(GTX125928, GenTex), rabbit polyclonal anti-IAV M2 (GTX125951, GenTex), rabbit polyclonal anti-IAV NS1(GTX125990, GenTex), rabbit polyclonal anti-IAV NS2 (GTX125953, GenTex), anti-FLAG M2 Affinity Gel (A2220, Sigma), anti-Strep-Tactin Sepharose (2-1201-010, IBA), mouse monoclonal anti-FLAG M2 (F3165, Sigma), mouse monoclonal Anti-Strep Tag (SAB2702216, Sigma), mouse monoclonal anti-Myc tag (AF0033, Beyotime), mouse monoclonal anti-GAPDH (HC301, Transgen), mouse monoclonal anti-Histone H2B (AH426, Beyotime), IRDye 800CW goat anti-rabbit IgG (926-32211, LC-COR), IRDye 800CW goat anti-mouse IgG (926-32210, LC-COR), FITC conjugated goat anti-mouse IgG (HS211, Transgen), Alexa Fluor 594 conjugated goat anti-rabbit IgG (H+L) (A-11012, Invitrogen).

Ren X., Yu Y., Li H., Huang J., Zhou A., Liu S., Hu P., Li B., Qi W, & Liao M. (2019). Avian Influenza A Virus Polymerase Recruits Cellular RNA Helicase eIF4A3 to Promote Viral mRNA Splicing and Spliced mRNA Nuclear Export. Frontiers in Microbiology, 10, 1625.

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Investigating Inflammasome Signaling Pathway

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RhS100A9, CLI-095, rhIL-1β and DPI were purchased from Abcam (ab95909), Invivogen (CA92121), Pepro Tech (200-01B) and Selleck (S8639), respectively. Primary antibodies were obtained from the following manufacturers: p53, p21, and p16 from Cell Signalling Biotechnology (Danvers, MA, USA); NLRP3 from Abcam (Cambridge, MA, USA); and GAPDH, Caspase-1, and IL-1β from Affinity Biosciences. Alexa Fluor 594-conjugated goat anti-rabbit IgG (H + L) and FITC conjugated anti-rabbit IgG (H + L) secondary antibodies were purchased from Thermo Scientific (Rockford, IL, USA) and Affinity Biosciences, respectively. HRP conjugated secondary antibodies were purchased from Affinity Biosciences.

Shi L., Zhao Y., Fei C., Guo J., Jia Y., Wu D., Wu L, & Chang C. (2019). Cellular senescence induced by S100A9 in mesenchymal stromal cells through NLRP3 inflammasome activation. Aging (Albany NY), 11(21), 9626-9642.

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Immunodetection of ADD1 and TPX2 Proteins

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All reagents used in this study were purchased from Sigma-Aldrich Co. (St. Louis, MO) unless otherwise noted. The rabbit polyclonal anti-ADD1 antibody (Catalog# NBP1-48,611) and the mouse monoclonal anti-TPX2 antibody (Catalog# NBP2-67,265) were purchased from Novus Biologicals. The rabbit polyclonal anti-p-ADD1 (S726) antibody (catalog# orb14892) was purchased from Biorbyt. Anti-α-tubulin-FITC (Catalog# F2168) and anti-γ-tubulin (Catalog# T6557) mouse monoclonal antibodies were obtained from Sigma-Aldrich. The mouse monoclonal anti-β-actin antibody (Catalog# ab49900) was purchased from Abcam. Alexa Fluor® 594-conjugated goat anti-rabbit IgG (H + L) (Catalog# A-11037) and Alexa Fluor® 647-conjugated goat anti-rabbit IgG (H + L) (Catalog# A-31633) were produced by Thermo Fisher Scientific Co.; TRITC conjugated goat anti-mouse IgG (H + L) (Catalog# ZF-0313) was purchased from Zhongshan Golden Bridge Biotechnology Co., LTD. Goat anti-rabbit IgG (Catalog# 1,706,515) and goat anti-mouse IgG (Catalog# 1,706,516) horseradish peroxidase (HRP)-conjugated antibodies were purchased from Bio-Rad.

Zhang Y., Fan B., Li X., Tang Y., Shao J., Liu L., Ren Y., Yang Y, & Xu B. (2022). Phosphorylation of adducin-1 by TPX2 promotes interpolar microtubule homeostasis and precise chromosome segregation in mouse oocytes. Cell & Bioscience, 12, 205.

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Immunohistochemical Staining of Brain Tissue

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Nissl staining solution (G1036) was purchased from Servicebio (Wuhan, China). Corn oil was purchased from Maclin Biochemical (Shanghai, China). CCT007093 was purchased from MedChemExpress (New Jersey, USA). Resveratrol was purchased from Aladdin (Shanghai, China). Optimal cutting temperature compound (OCT) was purchased from Sakura Finetek (Torrance, USA). Paraformaldehyde (PFA) was purchased from Biosharp (Hefei, China). Antibodies: Rabbit anti-Iba1 (ab178846) and Rabbit anti RBPMS (ab152101) were purchased from Abcam (Cambridge, UK). Mouse anti IL-1β (#12242) were purchased from Cell Signaling Technology (Boston, USA). Alexa Fluor 594–conjugated goat anti-rabbit IgG (H+L) and Alexa Fluor 488–conjugated goat anti-mouse IgG (H+L) were purchased from Thermo Fisher Scientific (Waltham, USA).

Mou Q., Yao K., Ye M., Zhao B., Hu Y., Lou X., Li H., Zhang H, & Zhao Y. (2021). Modulation of Sirt1-mTORC1 Pathway in Microglia Attenuates Retinal Ganglion Cell Loss After Optic Nerve Injury. Journal of Inflammation Research, 14, 6857-6869.

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Immunofluorescence Imaging of ERM Proteins

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The cells were processed in preparation for the Ab incubation, as described above. The cells were incubated overnight at 4 °C in the dark under humidified conditions with a rabbit anti-ezrin Ab at a dilution of 1:50, a rabbit anti-radixin Ab at a dilution of 1:50, or a rabbit anti-moesin Ab at a dilution of 1:25 in blocking buffer. After washing thrice with PBS-T, the cells were incubated for 60 min at room temperature with an Alexa Fluor 594-conjugated goat anti-rabbit IgG (H+L) (R37117; Thermo Fisher Scientific) prepared as dilution by adding by adding approximately 40 µL to 460 µL blocking buffer. Subsequently, the cells were washed thrice with PBS-T and incubated overnight at 4 °C in the dark under humidified conditions with an Alexa Fluor 488-conjugated rabbit anti-PD-L1 Ab (25048; Cell Signaling Technology). After washing thrice with PBS-T, Fluoro-KEEPER Antifade Regent Non-Hardening Type was added for storage and prevention of quenching. The preserved cells were observed and photographed at 0.5–0.7 µm intervals for z-axis at an original magnification of 60× with a Nikon Al confocal laser microscope system.

Kobori T., Tanaka C., Tameishi M., Urashima Y., Ito T, & Obata T. (2021). Role of Ezrin/Radixin/Moesin in the Surface Localization of Programmed Cell Death Ligand-1 in Human Colon Adenocarcinoma LS180 Cells. Pharmaceuticals, 14(9), 864.

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Immunofluorescence Staining of GLP-1R

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MC3T3-E1 cells were fixed in 4% paraformaldehyde, washed with 0.1% BSA in PBS, permeabilized with 0.3% Triton X-100. After blocking for 1 h, cells were incubated with anti-GLP-1R antibody (1:200) overnight at 4°C, followed by Alexa Fluor®594-conjugated goat anti-rabbit IgGH&L (Molecular Probes, USA) for 1 h. Images were observed under a FV1000 FLUOVIEW confocal laser scanning microscope (Olympus, Japan).

Wu X., Li S., Xue P, & Li Y. (2018). Liraglutide Inhibits the Apoptosis of MC3T3-E1 Cells Induced by Serum Deprivation through cAMP/PKA/β-Catenin and PI3K/AKT/GSK3β Signaling Pathways. Molecules and Cells, 41(3), 234-243.

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Immunofluorescent Analysis of Cytoskeletal Proteins in Oocytes

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Oocytes were fixed in PBS (pH 7.4) containing 4% paraformaldehyde for 30
min at room temperature. After the oocytes were washed in washing buffer (0.1%
tween-20 and 0.01% triton X-100 in PBS) twice, oocytes were permeabilized in
0.5% triton X-100 in PBS for 20 min at room temperature. After two washes, the
oocytes were blocked in washing buffer with 3% BSA for 1 h at room temperature
and then incubated overnight at 4°C with FITC-labelled mouse
anti-α-tubulin (1:200, Sigma-Aldrich) or mouse monoclonal
anti-γ-H2AX antibody, clone JBW301 (1:200, Millipore-Sigma) and
anti-phospho-p44/42 MAPK (Erk1/2) (1:200, Cell Signaling Technologies). As for
γ-H2AX or phospho-p44/42 MAPK (Erk1/2)staining, the oocytes were washed
in washing buffer for three times (10 min each time), then incubated with the
Alexa Fluor 488-conjugated goat anti-mouse IgG (H + L) and Alexa Fluor
594-conjugated goat anti-rabbit IgG (H + L) secondary antibodies (1:300,
Molecular Probes) for 1 h at 37°C, respectively. Then, oocytes were
washed another three times (10 min each time). Oocyte DNA was counterstained
with 1 μg/ml of 4′,6-diamidine-2′-phenylindole
dihydrochloride (DAPI) for 10 min at room temperature. After DAPI staining,
oocytes were mounted on glass slides with 80% glycerol, examined with a Nikon
A1R confocal microscope, and processed using NIS-Elements software.

Jiao X., Gonsioroski A., Flaws J.A, & Qiao H. (2020). Iodoacetic acid disrupts mouse oocyte maturation by inducing oxidative stress and spindle abnormalities. Environmental pollution (Barking, Essex : 1987), 268(Pt A), 115601.

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Fluorescent Probes for Cellular Assays

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Perfluorononanoic acid (PFNA, 97%), 3-Isobutyl-1-methylxanthine (IBMX, ≥ 99%) and 2′,7′-dichlorofluorescin diacetate (DCFH-DA, ≥ 97%) were obtained from Sigma-Aldrich (St. Louis, MO, USA). JC-1 Mitochondrial Membrane Potential Detection Kit and MitoView™ Green were purchased from Biotium (Hayward, CA, USA). Mouse monoclonal anti-α-Tubulin-FITC antibody and anti-phospho-Histone H2A.X (Ser139) antibody, clone JBW301 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-phospho-p44/42 MAPK (Erk1/2) antibody was purchased from Cell Signaling Technologies. Alexa Fluor 488-conjugated goat anti-mouse IgG (H + L) and Alexa Fluor 594-conjugated goat anti-rabbit IgG (H + L) secondary antibodies were obtained from Molecular Probes.

Jiao X., Liu N., Xu Y, & Qiao H. (2021). Perfluorononanoic acid impedes mouse oocyte maturation by inducing mitochondrial dysfunction and oxidative stress. Reproductive toxicology (Elmsford, N.Y.), 104, 58-67.

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