The largest database of trusted experimental protocols

Goat anti mouse igg h l fitc

Manufactured by Abcam
Sourced in United Kingdom

Goat anti-mouse IgG H&L (FITC) is a secondary antibody used for detection and visualization in immunoassays and other applications. The antibody is conjugated with the fluorescent dye FITC (Fluorescein isothiocyanate), which allows for fluorescent detection of target proteins or cells.

Automatically generated - may contain errors

13 protocols using goat anti mouse igg h l fitc

1

Immunostaining of Spinal Sections

Check if the same lab product or an alternative is used in the 5 most similar protocols
Frozen spinal sections (8 μm) were fixed with cold acetone for 10 min and
rinse with PBS for three times. The sections were then blocked with
10% normal donkey serum for 1 h at room temperature. Subsequently, the
sections were incubated with primary antibody, rabbit anti-GFAP
(1:100, 16825–1-AP) or rabbit anti-Iba1 (1:50, 10904–1-AP), overnight
at 4°C. After incubation and being washed with PBS, sections were
incubated with goat anti-mouse IgG H&L (FITC) (1:200, Abcam,
ab6785) at room temperature for 1 h and then examined with a
fluorescence microscope.
+ Open protocol
+ Expand
2

Immunofluorescence Staining of Mouse Skin

Check if the same lab product or an alternative is used in the 5 most similar protocols
Tissues were harvested from adult mice 6–8 weeks of age. Formalin fixed, paraffin embedded sections were processed by H&E staining for histologic evaluation by a board certified dermatopathologist blinded to the specimen genotype.
For immunofluorescence studies, tissue was embedded, frozen in OCT and cryosections prepared. Anti-mouse Dsg3 (clones AK18 and AK23, MBL, Woburn, MA) and anti-human Dsg3 (clone 5G11, AbD serotec, Kidlington, Oxford, UK) antibodies diluted in Tris-buffered-saline 5mM Ca++ (TBS/Ca++) were applied to the sections in a humidity chamber at RT for 30 minutes. Following three 10 minute washes, polyclonal goat anti-mouse IgG-H+L-FITC (abcam, Cambridge, MA) was used as a secondary antibody before evaluation under a fluorescence scope. Anti-mouse Dsc3 (gp2280, the kind gift of Dr. Peter Koch) was used to detect Dsc3 as above, but with a primary overnight incubation at 4°C followed by goat anti-guinea pig Alexa 488 (Life Technologies, Grand Island, NY). Similar studies were performed using patient sera followed by mouse anti-human IgG4 FITC (clone HP-6025, Sigma, St. Louis, MO) as a secondary antibody. For direct immunofluorescence studies, cryosections were directly stained with anti-human IgG4 FITC (clone HP-6025, Sigma, St. Louis, MO).
+ Open protocol
+ Expand
3

SGLT2 Expression in CaKi-1 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
We seeded 2×105 cells in a 6-well culture plate for 24-h culturing and incubated it with 2 μM dapagliflozin for 48 h. Then, the CaKi-1 cells were fixed with 4% paraformaldehyde for 20 min and with 0.2% Triton X-100 for 10 min, followed by blocking with 5% goat serum albumin. The cells were incubated with anti-SGLT2 antibody (1: 100, Abcam) in a wet box overnight at 4°C. Then, cells were incubated with Goat Anti-Mouse IgG H&L (FITC) (1: 1000, Abcam) for 1 h at 37°C and counterstained by DAPI (1 μg/mL) for 15 min. Images of the cells were obtained using a laser scanning confocal microscope (Leica, TCS SP5, Germany) and the fluorescent intensity was quantified by Image-Pro Plus 6.0 software.
+ Open protocol
+ Expand
4

Investigating Viral Protein Expression in BHK-21 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols
BHK-21 cells were seeded into a 6-well plate and cultured at 37 °C with 5% CO2. A cell monolayer at 80% confluence was transfected with either H, optiH, H-IRES-N, optiH-IRES-optiN, optiH-IRES-IFN, or an empty plasmid vector, pIRES, using Lipofectamine 2000 (Thermo Fisher, Carlsbad, CA 92010, USA) according to the manufacturer’s instructions. At 48 h after transfection, the cells were fixed in 1 mL of 4% formaldehyde for 30 min at 37 °C and permeabilized in 1 mL of 0.5% Triton X-100 for 10 min. After three washes with phosphate-buffered saline (PBS), samples were blocked with goat serum for 30 min at 37 °C. The samples were washed and blocked after each subsequent antibody exposure to anti-CDV H protein monoclonal antibody (1C42H11 monoclonal antibody, VMRD, Pullman, WA, 99163, USA), anti-CDV N protein monoclonal antibody (CDV-NP monoclonal antibody, VMRD, USA), and anti-mink IFN-γ monoclonal antibody [19 (link)], as appropriate for the corresponding plasmid vectors, whereas goat anti-mouse IgG H-L (FITC) (Abcam, Cambridge 02139, UK) was used as the secondary antibody. Images were obtained using a fluorescence microscope (EVOS FL, Thermo, USA). Five fields of vision for each image were randomly selected, and the mean number of fluorescent cells was calculated and compared to evaluate protein expression levels.
+ Open protocol
+ Expand
5

Immunofluorescence Staining of Mouse Skin

Check if the same lab product or an alternative is used in the 5 most similar protocols
Tissues were harvested from adult mice 6–8 weeks of age. Formalin fixed, paraffin embedded sections were processed by H&E staining for histologic evaluation by a board certified dermatopathologist blinded to the specimen genotype.
For immunofluorescence studies, tissue was embedded, frozen in OCT and cryosections prepared. Anti-mouse Dsg3 (clones AK18 and AK23, MBL, Woburn, MA) and anti-human Dsg3 (clone 5G11, AbD serotec, Kidlington, Oxford, UK) antibodies diluted in Tris-buffered-saline 5mM Ca++ (TBS/Ca++) were applied to the sections in a humidity chamber at RT for 30 minutes. Following three 10 minute washes, polyclonal goat anti-mouse IgG-H+L-FITC (abcam, Cambridge, MA) was used as a secondary antibody before evaluation under a fluorescence scope. Anti-mouse Dsc3 (gp2280, the kind gift of Dr. Peter Koch) was used to detect Dsc3 as above, but with a primary overnight incubation at 4°C followed by goat anti-guinea pig Alexa 488 (Life Technologies, Grand Island, NY). Similar studies were performed using patient sera followed by mouse anti-human IgG4 FITC (clone HP-6025, Sigma, St. Louis, MO) as a secondary antibody. For direct immunofluorescence studies, cryosections were directly stained with anti-human IgG4 FITC (clone HP-6025, Sigma, St. Louis, MO).
+ Open protocol
+ Expand
6

Anti-IL-37 Monoclonal Antibody Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
Anti-IL-37 monoclonal antibody, Goat Anti-Mouse IgG H&L (FITC), and Mouse IgG1 were purchased from Abcam Corporation (Cambridge, UK). TWG (10 mg/tablet, Approval number Z35020431) was obtained in the form of tablets (Tripterygium wilfordii glycoside tablets) from Fujian Huitian Bio-pharma Co., Ltd. (GMP certificated, Fujian, China). The tablets were dissolved in the incubation medium at a final concentration of 5 mg mL−1 as stock solution. Inhibitor of extracellular signal-regulated kinase 1 (ERK1)/ERK2, U0126, and inhibitor of p38 MAPK, SB 203580, were from Cell Signaling Technology (Beverly, MA, USA). RNAiso Plus, Prime Script™ RT Master Mix and SYBR® Premix Ex Taq (Perfect Real Time) were from Takara Biotechnology Co., Ltd. (Dalian, China). Primers were synthesized by Sangon Biotech Co., Ltd. (Shanghai, China). eBioscience Fixation/Permeabilization Concentrate and Permeabilization Buffer (10×) were from eBioscience Inc. (San Diego, CA, USA). Dimethyl sulfoxide (DMSO) and phorbol-12-myristate 13-acetate (PMA) were from Sigma (St. Louis. MO, USA). THP-1 cell line was obtained from the Cell bank of Chinese Academy of Sciences, Shanghai, China. Cells culture reagents including 1640 medium and fetal bovine serum (FBS) were purchased from HyClone (Longan, UT, USA).
+ Open protocol
+ Expand
7

Pericentrin Expression Analysis in PBMCs

Check if the same lab product or an alternative is used in the 5 most similar protocols
Flow cytometry was used to examine the expression of PCNT protein in peripheral blood mononuclear cells (PBMCs) of the patient and her parents. PBMCs were collected from peripheral blood of the above subjects using lymphocyte separation medium (TBD Science, LTS1077). Then, the separated PBMCs were washed with phosphate‐buffered saline (PBS) and centrifuged (4°C, 300 g, 10 min). Cell fixation and permeabilization were performed successively using the perfix‐nc Kit (Beckman Coulter, A07803) according to the manufacturer's instructions. Next, 1:200 diluted mouse anti‐pericentrin antibody (Abcam, ab28144, UK) was added and incubated at room temperature for 30 min. After centrifugation (4°C, 360 g, 5 min), the supernatant was discarded while the precipitate was washed again and resuspended in 1ml PBS (precooling at 4°C). Finally, 1:400 diluted goat anti‐mouse IgG H&L (FITC; Abcam, ab6785) was added for staining in the dark. BD FACSCanto II flow cytometer, FACSDiva, and FlowJo v10 software were used to collect and analyze data.
+ Open protocol
+ Expand
8

Immunofluorescence Assay for PEDV Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols
After infection with PEDV strains, the obvious cytopathic effect (CPE) of Vero E6 cells was observed at 48 h. Then, the cells were fixed with 4% paraformaldehyde at room temperature for 30 min, followed by blocking with 5% BSA at 37 °C for 1 h. Each hybridoma supernatant was added and incubated overnight at 4 °C. After three washes, the cells were stained with goat anti-mouse IgG H&L (FITC) (1:1000 dilution) (Abcam, UK), followed by incubation for 1 h at 37 °C. Images of the stained cells were then visualized using an inverted fluorescence microscope (DMi8, Leica Microsystems).
+ Open protocol
+ Expand
9

Immunofluorescence Analysis of Lin28a, α-SMA, and C/EBP in Paraffin-Embedded Tissue

Check if the same lab product or an alternative is used in the 5 most similar protocols
Paraffin-embedded sections (4 μm) were de-paraffinized with xylene and rehydrated with gradient concentrations of alcohol. The sections were then placed in 0.01 mol/L citrate buffer for high-pressure repair. Endogenous peroxidase activity was blocked using H 2 O 2 . Sections were incubated overnight with rabbit anti-Lin28a (1:200, Proteintech, Wuhan, China), mouse anti-α-SMA (1:200, Cell Signalling Technology, Danvers, MA, USA), mouse anti-C/EBP (1:200, Proteintech), and rabbit anti-α-SMA (1:200, Cell Signalling Technology) at 4°C. After washing with phosphate buffered saline (PBS), the sections were incubated with goat anti-mouse IgG H&L (FITC) (1:500, Abcam, Cambridge, UK) or goat anti-rabbit IgG H&L (Alexa Fluor ® 647) (1:500, Abcam) for 1 h. Finally, the nuclei were stained with 4',6-diamidino-2-phenylindole (DAPI; Solarbio) for 5 min.
+ Open protocol
+ Expand
10

Bovine Viral Diarrhea Virus Antibody Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
BVDV−1 E2 specific monoclonal antibody (mAb) stored in the laboratory. The BVDV FITC conjugate was purchased from VMRD (CJ-F-BVD-10 mL). BVDV−1 mAb E2 gp53 IgG2a Isotype was purchased from VMRD (CATALOG #: 157). BoHV−1 positive serum was stored in the laboratory. Goat Anti-Mouse IgG H&L (FITC) and Goat Anti-Mouse IgG H&L (HRP) were purchased from abcam (ab6785, ab6789). MONTANIDETM ISA 206 VG adjuvant was purchased from Seppic (Shanghai, China). Propolis adjuvant was purchased from Jinhe-Youben (Hohhot, China).
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!