The DRG at the L5 spinal level was collected 5 h and 8 days after i.p. oxaliplatin at 5 mg/kg, fixed in 4% paraformaldehyde in 0.1 M phosphate buffer, and subjected to immunofluorescence staining using a chicken anti-HMGB1 polyclonal antibody (#326059669, SHINO-TEST Corporation, Tokyo, Japan) (1:1000 dilution), Alexa Fluor 568-conjugated anti-chicken IgY (H + L) antibody (A-11041, Thermo Fisher Scientific Corporation, Carlsbad, CA, USA) (1:1000 dilution), as mentioned above.
Chicken anti hmgb1 polyclonal antibody
Manufactured by Shino-Test
Sourced in Japan
Chicken anti-HMGB1 polyclonal antibody is a laboratory reagent used to detect and quantify the presence of High Mobility Group Box 1 (HMGB1) protein in biological samples. HMGB1 is a nuclear protein involved in various cellular processes. This antibody can be used in techniques such as Western blotting, ELISA, and immunohistochemistry to identify and measure HMGB1 levels.
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Lab products found in correlation
Puromycin dihydrochloride, by Merck Group (1 mentions)
Tak 242, by Cayman Chemical (1 mentions)
Anti rage antibody, by Santa Cruz Biotechnology (1 mentions)
Horseradish peroxidase hrp conjugated igg antibody, by Cell Signaling Technology (1 mentions)
Hrp conjugated igg antibody, by Cell Signaling Technology (1 mentions)
Alexa fluor 488 conjugated igg h l f ab 2 fragment, by Cell Signaling Technology (1 mentions)
Anti cgrp antibody, by Abcam (1 mentions)
Hmgb1 antibody, by GeneTex (1 mentions)
Anti p44 42 mapk antibody, by Cell Signaling Technology (1 mentions)
Anti tlr4 antibody, by Cell Signaling Technology (1 mentions)
Fps zm1, by Cayman Chemical (1 mentions)
Anti β actin antibody, by Abcam (1 mentions)
C57bl 6j b6 mice, by Jackson ImmunoResearch (1 mentions)
4 protocols using chicken anti hmgb1 polyclonal antibody
1
Oxaliplatin-Induced HMGB1 Expression in L5 DRG
2
Investigating HMGB1 and RAGE Signaling
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Puromycin dihydrochloride (#P9620) was purchased from Sigma-Aldrich; Merck KGaA. TLR4 antagonist TAK-242 (#13871) and RAGE antagonist FPS-ZM1 (#11909) were purchased from Cayman Chemical Co. Chicken anti-HMGB1 polyclonal antibody (#326052233) was purchased from Shino-Test. Control shRNA plasmid-A (#sc-108060), HMG-1 shRNA plasmid (#sc-37982-SH), and anti-RAGE antibody (anti-mouse, monoclonal, sc-80652) were purchased from Santa Cruz Biotechnology, Inc. HMGB1 antibody (anti-mouse, monoclonal, GTX628834) was purchased from GeneTex, Inc. Anti-phospho-p44/42 MAPK antibody (p-ERK; anti-rabbit, monoclonal, #4370), anti-p44/42 MAPK antibody (ERK; anti-rabbit, monoclonal, #4695), horseradish peroxidase (HRP)-conjugated IgG antibody (goat anti-rabbit, monoclonal, #7074), HRP-conjugated IgG antibody (goat anti-mouse, monoclonal, #7076), and Alexa Fluor 488-conjugated IgG (H+L) F(ab′)2 fragment (goat anti-rabbit, monoclonal, #4412) were purchased from Cell Signaling Technology, Inc. Anti-TLR4 antibody (anti-rabbit, polyclonal, #ab13556), anti-β-actin antibody (anti-mouse, monoclonal, #ab49900), anti-CGRP antibody (anti-goat, polyclonal, #ab36001), and Alexa Fluor 647-conjugated IgG H&L antibody (donkey anti-goat, monoclonal, #ab150135) were purchased from Abcam.
3
Murine Autoimmune Uveitis Model
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Eight- to ten-week-old female C57BL/6J (B6) mice, purchased from the Jackson Laboratory (Bar Harbor, ME, USA), were housed and maintained in the animal facilities of the University of Louisville (KY, USA). All animal studies conformed to the Association for Research in Vision and Ophthalmology statement about the use of animals in ophthalmic and vision research. The protocol (#14052) was approved by the Institutional Animal Care and Use Committee of the University of Louisville.
All T cells were cultured in complete medium [RPMI 1640 medium (Mediatech, Manassas, VA, USA) supplemented with 10% fetal calf serum (Hyclone, Logan, UT, USA), 5 × 10−5 M 2-mercapatoethanol, and 100 µg/ml of penicillin/streptomycin]. The human IRBP peptide 1–20 (GPTHLFQPSLVLDMAKVLLD) was synthesized by Sigma-Aldrich (St. Louis, MO, USA). The fully reduced HMGB1 (Cat#HM-115) was purchased from HMGBiotech (Milano, Italy) and chicken anti-HMGB1 polyclonal antibody (Cat#326052233) from Shino-Test Corporation (Kanagawa, Japan).
All T cells were cultured in complete medium [RPMI 1640 medium (Mediatech, Manassas, VA, USA) supplemented with 10% fetal calf serum (Hyclone, Logan, UT, USA), 5 × 10−5 M 2-mercapatoethanol, and 100 µg/ml of penicillin/streptomycin]. The human IRBP peptide 1–20 (GPTHLFQPSLVLDMAKVLLD) was synthesized by Sigma-Aldrich (St. Louis, MO, USA). The fully reduced HMGB1 (Cat#HM-115) was purchased from HMGBiotech (Milano, Italy) and chicken anti-HMGB1 polyclonal antibody (Cat#326052233) from Shino-Test Corporation (Kanagawa, Japan).
4
Anti-HMGB1 Antibody Microinjection in Rat PVN
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Rats received bilateral PVN microinjections of chicken anti-HMGB1 polyclonal antibody (Shino-Test Corporation, Tokyo, Japan; 10 μg in 10 μL of 10 mM of Tris-buffered saline, pH 7.4, 5 μL every second day for seven consecutive days), a dose of anti-HMGB1 polyclonal Ab similar to that used in previous studies, or an equivalent volume of control IgG. The PE pipe with a length of 15–20 cm and the casing pipe with a diameter of 0.4 mm were connected, and these were subsequently connected to a microsyringe. The bilateral PVN microinjections were carried out using a micropump injection device (RWD Life Science Co., Shenzhen, China). The bilateral PVN microinjection volume was 5 μL for each site. The injection rate was 0.5 μL/min. After the injection, the syringe was left for an additional 5 min before it was slowly retracted. At the end of the experiment, 50 nL of Evans Blue (2%) was injected into the microinjection site for histological identification. Only the data from rats, in which the microinjection sites were within the boundaries of the PVN were used for analysis. Rats with microinjection sites outside the PVN or at the margin of the PVN were excluded from the data analysis.
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