Fresh-frozen skin samples were used for immunohistochemistry. We used the following antibodies as primary antibodies: a mouse monoclonal antibody directed against CXCR3 (1C6, BD Biosciences Pharmingen, Heidelberg, Germany), a rabbit antibody against CCR3 (MBL International, Woburn, MA, USA), a mouse monoclonal antibody directed against CCR4 (1G1, BD Biosciences Pharmingen) and a goat antibody against CCR10 www.medicaljournals.se/acta (Capralogics, Hardwick, MA, USA). The following antibodies were used as secondary ones: a biotinylated horse anti-mouse IgG antibody (Vector Laboratories, Burlingame, CA, USA) for CXCR3 and CCR4, a biotinylated goat anti-rabbit IgG antibody (Vector Laboratories) for CCR3 and a biotinylated rabbit anti-goat IgG antibody (Vector Laboratories) for CCR10. We classified the immunostaining intensity into 3 categories: negative (-), weakly positive (+) and strongly positive (++). Samples in which more than half of the infiltrating cells were positive for a chemokine receptor were considered strongly positive for that receptor. Four doctors evaluated the staining blinded to the clinical diagnosis.
Biotinylated goat anti rabbit igg antibody
Manufactured by Vector Laboratories
Sourced in United States
The Biotinylated goat anti-rabbit IgG antibody is a secondary antibody that binds to rabbit immunoglobulin G (IgG) molecules. It is labeled with biotin, which can be detected using various detection systems.
Automatically generated - may contain errors
Lab products found in correlation
Ba 1000, by Vector Laboratories (3 mentions)
Histovt one, by Nacalai Tesque (3 mentions)
Biotinylated horse anti mouse igg antibody, by Vector Laboratories (3 mentions)
Goat serum, by Cell Signaling Technology (2 mentions)
Signalstain dab substrate kit, by Cell Signaling Technology (2 mentions)
Vectastain abc kit, by Vector Laboratories (2 mentions)
Vectastain elite abc kit, by Vector Laboratories (2 mentions)
Lsm 780, by Zeiss (2 mentions)
Vectastain elite abc standard kit, by Vector Laboratories (2 mentions)
Aec substrate kit, by Vector Laboratories (2 mentions)
Streptavidin peroxidase conjugate, by Jackson ImmunoResearch (2 mentions)
Rabbit anti cleaved caspase 9, by Cell Signaling Technology (2 mentions)
Anti id1, by Abcam (1 mentions)
α sma, by Abcam (1 mentions)
Scanscope, by Leica Biosystems (1 mentions)
Imagescope, by Leica Biosystems (1 mentions)
Vectastain abc ap staining kit, by Vector Laboratories (1 mentions)
Rabbit anti tyrosine hydroxylase th, by Merck Group (1 mentions)
Rabbit anti nurr1, by Santa Cruz Biotechnology (1 mentions)
Rabbit anti gdnf, by Santa Cruz Biotechnology (1 mentions)
37 protocols using biotinylated goat anti rabbit igg antibody
1
Immunohistochemical Analysis of Chemokine Receptors
2
Immunohistochemical Analysis of ESCC Tissues
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A total of 36 pairs of ESCC tissues and their adjacent non-cancer tissues were obtained from the Tissue Bank of the Laboratory for Cancer Signal Transduction, Xinxiang Medical University (Xinxiang, China). All procedures were approved by the Institutional Review Board of Xinxiang Medical University.
Immunohistochemical staining was conducted as previously described (23 (link)). In brief, formalin-fixed, paraffin-embedded tissues were sectioned and deparaffinized with xylene, rehydrated with gradient ethanol and distilled water, and were subsequently blocked with serum and incubated with anti-AFT3 (1:100, ca no. ab191513) and anti-ID1 (1:100, cat no. ab134163) primary antibodies (Abcam, Cambridge, MA, USA) at 4°C overnight. The sections were then incubated with a biotinylated goat anti-rabbit IgG antibody, dilution (1:200; cat no. PI-1000; Vector Laboratories, Inc., Burlingame, CA, USA), and a VECTASTAIN Elite ABC kit (Vector Laboratories, Inc., Burlingame, CA, USA) was used according to the manufacturer's protocol. The sections were finally stained with 3,3′-diaminobenzidine. Immunohistochemical staining was evaluated based on immunostaining intensities (absent or weak, moderate and strong) as previously described (23 (link)).
Immunohistochemical staining was conducted as previously described (23 (link)). In brief, formalin-fixed, paraffin-embedded tissues were sectioned and deparaffinized with xylene, rehydrated with gradient ethanol and distilled water, and were subsequently blocked with serum and incubated with anti-AFT3 (1:100, ca no. ab191513) and anti-ID1 (1:100, cat no. ab134163) primary antibodies (Abcam, Cambridge, MA, USA) at 4°C overnight. The sections were then incubated with a biotinylated goat anti-rabbit IgG antibody, dilution (1:200; cat no. PI-1000; Vector Laboratories, Inc., Burlingame, CA, USA), and a VECTASTAIN Elite ABC kit (Vector Laboratories, Inc., Burlingame, CA, USA) was used according to the manufacturer's protocol. The sections were finally stained with 3,3′-diaminobenzidine. Immunohistochemical staining was evaluated based on immunostaining intensities (absent or weak, moderate and strong) as previously described (23 (link)).
3
Lung Tissue Immunohistochemistry: CD31 and α-SMA Analysis
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Infected lung tissues were fixed and sectioned by the Histology lab of Animal Health Diagnostic Center in Cornell University. Briefly, unstained slides were hydrated and stained with primary antibodies (CD31, 1:1000, BD; α-SMA, 1:500, Abcam) overnight at 4°C. Slides were washed and stained with secondary antibodies (1:200, biotinylated goat anti-rabbit IgG antibody, Vector lab) and ABC kit at room temperature for 2h. Sections were developed with DAB and mounted for digital scanning (Scanscope, Aperio Technologies). Blind quantitative analysis of IHC images were performed with the Imagescope (Aperio Technologies) software.
4
ILDR2 Expression in Inflammatory Bowel Disease
5
Producing Rabbit Polyclonal Antibody Against PRV
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For producing rabbit polyclonal antibody against PRV, rabbits were injected with three doses of Geskypur PRV vaccine (Merial Animal Health, Woking, UK) 20 days apart. Twenty days after the last injection, rabbits were exsanguinated, and the collected blood serum was titrated by anti-PRV-gB ELISA and stored at −20 °C. The biotinylated goat anti-rabbit IgG antibody was purchased from Vector Laboratories (Burlingame, CA, USA). Goat normal serum was used as blocking serum.
For the inoculations we used the well-studied wild-type Kaplan (Ka) strain, which is virulent for newborn pigs (Kritas, 1994c). The Ka strain was kindly provided by the Laboratory of Virology, Faculty of Veterinary Medicine, University of Ghent.
For the inoculations we used the well-studied wild-type Kaplan (Ka) strain, which is virulent for newborn pigs (Kritas, 1994c). The Ka strain was kindly provided by the Laboratory of Virology, Faculty of Veterinary Medicine, University of Ghent.
6
Immunohistochemical Analysis of Dopaminergic Neurons
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Animals were sacrificed via pentobarbital overdose (60 mg/kg) and intracardially perfused with saline, then 4% paraformaldehyde. Brains were removed and post-fixed in 4% paraformaldehyde for 24 h and then dehydrated in 30% sucrose. The sections at a thickness of 16 μm were cut through the substantia nigra (−2.92 to −3.88 mm from bregma) (Paxinos and Watson, 2007 ). The tissue sections were incubated in 0.3% H2O2 for 45 min, and blocked in 10% goat serum for 1 h. The sections were incubated overnight at 4°C with rabbit anti-tyrosine hydroxylase (TH; 1:300; Millipore), rabbit anti-dopamine transporter (DAT; 1:200; Santa Cruz), rabbit anti-GDNF (1:200; Santa Cruz Biotechnology) or rabbit anti-Nurr1 (1:100; Santa Cruz Biotechnology). Subsequently, the sections were incubated with biotinylated goat anti-rabbit IgG antibody (1:250; Vector labs) and avidin-biotin complex (ABC-kit, Vector Labs) at 37°C for 30 min, and then stained with 3,3′-diaminobenzidine (DAB). Then the sections were observed under a light microscope (BX51; Olympus, Tokyo, Japan). Image scanning analysis system (Image-Pro Plus) was used to analyze the changes in integrated optical density (IOD) of DAT, TH and GFAP.
7
Immunohistochemical Analysis of IL-33 and mMCP-8 in Excised Eye Tissues
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Excised eye specimens were fixed with 4% (wt/vol) paraformaldehyde and then embedded in paraffin. The tissues were sectioned at 4-μm thickness, and deparaffinized sections were subjected to hematoxylin and eosin (H&E) staining or immunohistochemistry. Immunofluorescence for IL-33 was as described previously7 (link). In brief, sections were incubated with an affinity-purified rabbit anti-mouse IL-33 polyclonal antibody, and bound antibodies were visualized with a biotinylated goat anti-rabbit IgG antibody (Vector Laboratories) and a Streptavidin, Alexa Fluor® 594 conjugate (Invitrogen, Carlsbad, CA). Immunofluorescence for mMCP-8 was as described previously9 (link). Sections were incubated with anti-mMCP-8 antibody (clone TUG8, BioLegend), and bound antibodies were visualized with a Cy3-conjugated donkey anti-rat IgG antibody (WAKO, Osaka, Japan). Following mounting with a ProLong® Diamond Antifade with DAPI (Life Technologies, Gaithersburg, MD), fluorescence images were recorded using a confocal laser scanning microscope LSM780 (Carl Zeiss MicroImaging, Thornwood, NY).
8
Tyrosine Hydroxylase Quantification in Brain Slices
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Brain slices obtained for the c-Fos experiments in basal conditions were used for TH quantification (n = 8 and 5 for control and MPH rats, respectively). Free-floating sections were incubated for 24 hours at 4°C with rabbit polyclonal anti-TH antibody (1:500; AB152, Santa Millipore), and for 2 hours at room temperature with a biotinylated goat anti-rabbit IgG antibody (1:400; Vector Laboratories). Tissue sections were further processed using the ABC Vectastain Elite kit (Vector Laboratories) and 3.3’-diaminobenzidine detection. The analyses of TH immunohistochemistry are detailed the Supplementary Materials section .
9
Immunohistochemical Detection of Factor VIII
10
Antibody Panel for Iron Homeostasis and Macrophage Markers
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Anti-LXRa rabbit polyclonal(USBiological Cat # 364027), Anti-LXRb rabbit polyclonal (Abcam Cat # ab28479), Anti-FTH1 rabbit polyclonal (Cell Signaling Cat # 3998 s), Anti-SLC40A1 rabbit polyclonal (Novus Biological Cat # NBP1-21502SS), Anti-TFR1 rabbit polyclonal (Abcam Cat # ab84036), Anti-ARG2 rabbit polyclonal (ThermoFischer Cat # PA5-27987), Anti-SLC7A2 rabbit polyclonal (ThermoFischer Cat # PA5-77552), Primary- Anti F4/80 Rat Monoclonal (Abcam Cat # ab6640), Secondary- Biotinylated rabbit anti-Rat IgG (Vector Laboratories Cat # BA-4001), Mouse IgG HRP linked whole antibody (GE Healthcare Cat # NA931V), Rabbit IgG HRP linked whole antibody (GE healthcare Cat # NA934V), Anti-F4/80 antibody [CI:A3-1] (AbcamCat # ab6640), Anti-TRF1 antibody (GeneTex Cat # GTX102596), Peroxidase AffiniPure F(ab’)2 Fragment Donkey Anti- Rabbit IgG (H + L) (Jackson ImmunoResearch Cat # 711-036-152), Anti-SLC40A1 antibody (Novus Biological Cat # NBP1-21502), Biotinylated Goat Anti-Rabbit IgG Antibody (Vector Laboratories Cat # BA-1000).
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