The rabbit monoclonal antibodies (mAb) against phospho-eIF2α (Ser51) and eIF2α were purchased from Cell Signaling Technology (Beverly, MA, USA). Mouse mAb against beta-actin was obtained from Medical and Biological Laboratories (MBL, Nagova, Japan). Antibodies against TIA1 (goat polyclonal), G3BP1 (rabbit polyclonal), TIAR (goat polyclonal), and eIF3η (goat polyclonal) were purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, CA, USA). Rabbit mAb against NF-κB p65 and phospho-NF-κB p65 were purchased from Cell Signaling Technology. Mouse mAb against PRRSV nucleocapsid (N) protein was prepared by our laboratory. Alexa Fluor 594-conjugated donkey anti-goat IgG, Alexa Fluor 594-conjugated donkey anti-rabbit IgG, Alexa Fluor 488-conjugated donkey anti-rabbit IgG, Alexa Fluor 488-conjugated donkey anti-mouse IgG, and Alexa Fluor 647-conjugated donkey anti-mouse IgG were also obtained from Santa Cruz Biotechnology, Inc. Horseradish peroxidase (HRP)-conjugated goat anti-rabbit, HRP-conjugated goat anti-mouse, and HRP-conjugated donkey anti-goat were purchased from Medical and Biological Laboratories (MBL).
Alexa fluor 488 conjugated donkey anti mouse igg
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Alexa Fluor 488-conjugated donkey anti-mouse IgG is a secondary antibody that binds to mouse immunoglobulin G (IgG) and is labeled with the Alexa Fluor 488 fluorescent dye. It can be used for the detection and visualization of mouse primary antibodies in various immunoassays and imaging applications.
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Lab products found in correlation
Alexa fluor 594 conjugated donkey anti rabbit igg, by Santa Cruz Biotechnology (3 mentions)
Alexa fluor 488 conjugated donkey anti rabbit igg, by Santa Cruz Biotechnology (1 mentions)
Phospho nf κb p65, by Cell Signaling Technology (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Horseradish peroxidase conjugated goat anti rabbit antibody, by Beyotime (1 mentions)
Bhk 21, by Thermo Fisher Scientific (1 mentions)
Horseradish peroxidase conjugated goat anti mouse antibody, by Beyotime (1 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Llc pk1 cells, by American Type Culture Collection (1 mentions)
Bhk 21 cells, by National Collection of Authenticated Cell Cultures (1 mentions)
Hela cells, by National Collection of Authenticated Cell Cultures (1 mentions)
Confocal microscopy, by Zeiss (1 mentions)
Rabbit anti cd31, by Abcam (1 mentions)
Lsm 510, by Zeiss (1 mentions)
Calci clear rapid, by National Diagnostics (1 mentions)
Alexa fluor 594 conjugated donkey anti mouse igg, by Santa Cruz Biotechnology (1 mentions)
Anti β actin antibody, by Beyotime (1 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (1 mentions)
4 protocols using alexa fluor 488 conjugated donkey anti mouse igg
1
Antibody Characterization for Protein Detection
2
Cell Culture and Virus Isolation Protocol
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IPI-2I cells (porcine intestinal epithelial cells), BHK-21 cells, and HeLa cells were obtained from the China Center for Type Culture Collection (Wuhan, China). LLC-PK1 cells were acquired from the American Type Culture Collection (ATCC CL-101; Manassas, VA, USA). IPI-2I, BHK-21, and LLC-PK1 cells were cultured in Dulbecco’s modified Eagle medium with 10% fetal bovine serum (Invitrogen, USA). HeLa cells were grown in RPMI 1640 medium with 10% fetal bovine serum. These cells were maintained in 5% CO2 at 37 °C. PDCoV strain CHN-HN-2014 (GenBank accession no. KT336560)10 (link) and TGEV strain WH1 (GenBank accession no. HQ462571) were isolated in 2014 and 2010, respectively, in China. Mouse monoclonal antibodies against TGEV M, TGEV N or PDCoV S, PDCoV N were created in-house. APN antibody was purchased from ABclonal (China). An anti-Flag rabbit polyclonal antibody (MBL, Japan), Alexa Fluor 594-conjugated donkey anti-rabbit IgG (Santa Cruz, USA), and Alexa Fluor 488-conjugated donkey anti-mouse IgG (Santa Cruz, USA) were used for indirect IFA. Horseradish peroxidase-conjugated goat anti-mouse antibody (Beyotime, China) and horseradish peroxidase-conjugated goat anti-rabbit antibody (Beyotime, China) were applied in western blots.
3
Immunohistochemical Assessment of Vascular Formation
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All specimens were decalcified in rapid decalcifying solution (Calci-Clear Rapid, National Diagnostics, Atlanta, GA) for 10 days and then embedded in paraffin, and cut into 7 μm sections. The sections were deparaffinized in xylene, rehydrated with a graded series of alcohols, and stained with hematoxylin and eosin solution.
Immunofluorescence assay was performed to evaluate vascular formation and pericyte recruitment. Briefly, histology sections were deparaffinized, rehydrated, and incubated in phosphate buffered saline solution with Tween-20 (PBS-T), containing 5% normal serum for 2 hrs. After overnight incubation with primary mouse anti-NG2 (1:100; Abcam, Cambridge, UK) and rabbit anti-CD31 (1:50; Abcam, Cambridge, UK) antibodies, the histology sections were treated with Alexa Fluor 488-conjugated donkey anti-mouse IgG and Alexa Fluor 555-conjugated goat anti-rabbit IgG (Santa Cruz Biotechnology, Delaware, CA, USA). Cell nuclei were stained with 4′, 6-Diamidino-2-phenylindole dihydrochloride (DAPI). Immunoreactivity was detected by confocal microscopy (Carl Zeiss, Jena, Germany). The fluorescence intensity was measured from 4 different samples (at least 5 foci per section) by image analyzer LSM 510 software (Carl Zeiss, Jena, Germany).
Immunofluorescence assay was performed to evaluate vascular formation and pericyte recruitment. Briefly, histology sections were deparaffinized, rehydrated, and incubated in phosphate buffered saline solution with Tween-20 (PBS-T), containing 5% normal serum for 2 hrs. After overnight incubation with primary mouse anti-NG2 (1:100; Abcam, Cambridge, UK) and rabbit anti-CD31 (1:50; Abcam, Cambridge, UK) antibodies, the histology sections were treated with Alexa Fluor 488-conjugated donkey anti-mouse IgG and Alexa Fluor 555-conjugated goat anti-rabbit IgG (Santa Cruz Biotechnology, Delaware, CA, USA). Cell nuclei were stained with 4′, 6-Diamidino-2-phenylindole dihydrochloride (DAPI). Immunoreactivity was detected by confocal microscopy (Carl Zeiss, Jena, Germany). The fluorescence intensity was measured from 4 different samples (at least 5 foci per section) by image analyzer LSM 510 software (Carl Zeiss, Jena, Germany).
4
TGEV Infection Signaling Pathway
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Porcine kidney (PK-15) cells and HEK-293T cells were cultured in Dulbecco’s modified Eagle’s medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum in a humidified incubator with 37°C/5% CO2. TGEV strain WH-1 (GenBank accession no. HQ462571) was propagated and titered in PK-15 cells. Recombinant VSV-expressing green fluorescent protein (VSV-GFP) was generously provided by Prof. Zhigao Bu from the Harbin Veterinary Research Institute, China. Rabbit polyclonal antibodies against p65, IRF3, phosphorylated IRF3 (p-IRF3), and DDX1 were purchased from ABclone (Wuhan, China). Rabbit polyclonal antibody against phosphorylated p65 (p-p65) was purchased from Cell Signaling Technology (Beverly, MA, USA). Anti-β-actin antibody was purchased from Beyotime (Nantong, China). Mouse monoclonal antibodies (MAbs) against hemagglutinin (HA) and Flag were purchased from Medical and Biological Laboratories (MBL, Nagoya, Japan). MAb against TGEV N protein was prepared by our laboratory. Horseradish peroxidase (HRP)-conjugated goat anti-rabbit antibody and HRP-conjugated goat anti-mouse antibody were purchased from MBL. Alexa Fluor 594-conjugated donkey anti-rabbit IgG, Alexa Fluor 594-conjugated donkey anti-mouse IgG, and Alexa Fluor 488-conjugated donkey anti-mouse IgG were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA).
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