Hta125

Manufactured by Thermo Fisher Scientific

Sourced in United States

The HTA125 is a compact and efficient heating thermal analyzer designed for thermal analysis applications. It provides precise temperature control and measurement capabilities for a variety of sample types. The core function of the HTA125 is to accurately measure and analyze the thermal properties of materials.

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Lab products found in correlation

Gd2 f4, by Thermo Fisher Scientific (3 mentions) Tlr6.127, by Abcam (3 mentions) Elisa kit for von willebrand factor, by Nanjing Jiancheng (3 mentions) Histone h4, by Merck Group (3 mentions) Myeloperoxidase mpo detection kit, by Nanjing Jiancheng (3 mentions) Ac h4 acetyl k5 k8 k12 k16, by Abcam (2 mentions) Biotin lps, by InvivoGen (1 mentions) Accuri c6 flow cytometer, by BD (1 mentions) Ab254360, by Abcam (1 mentions) Ab181603, by Abcam (1 mentions) Ab288151, by Abcam (1 mentions) Ab207297, by Abcam (1 mentions) Ab76302, by Abcam (1 mentions) Polyvinylidene difluoride membrane, by Merck Group (1 mentions) Tnf α quantikine elisa kit, by R&D Systems (1 mentions) Carboxylate modified fluorescent microspheres, by Thermo Fisher Scientific (1 mentions)

6 protocols using hta125

Quantifying TLR4 Binding via Yeast Display

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Yeast cells with the Aga2p fusion were induced. After 10 minutes of vigorous vortexing, 100nM biotin-LPS (biotinylated ultra-pure LPS-EB, InvivoGen; LPS concentration was based on a formula weight of biotin-LPS of approximately 10,000) was incubated at a 1:3 molar ratio with human CD14 in a 37°C water bath for one hour. biotin-LPS/CD14 was then added at various concentrations to 1×10⁶ yeast cells and incubated at 37°C for one hour. Cells were washed and incubated with SA-Alexafluor647 (SA647). After washing, cells were analyzed on an Accuri C6 flow cytometer.
To detect TLR4 binding, 1×10⁶ yeast cells were incubated with various concentrations of flag-tagged TLR4, and after washing with PBS+0.5%BSA, the cells were incubated with anti-human TLR4 mAb HTA-125 at a 1:200 dilution (eBioscience). After one hour on ice, cells were washed and incubated with GaM647. After washing, cells were analyzed on an Accuri C6 flow cytometer. A negative control included yeast displayed protein L3, which is a Vβ domain with approximately the same size as MD-2 (Mattis et al., 2013 ).

Mattis D.M., Chervin A., Ranoa D., Kelley S., Tapping R, & Kranz D.M. (2015). Studies of the TLR4-associated protein MD-2 using yeast-display and mutational analyses. Molecular immunology, 68(2 0 0), 203-212.

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Western Blot Analysis of Inflammatory Signaling

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After lysing samples with RIPA lysis buffer (SY4680, YITA), the concentration of protein was determined using a Dxy_PIERCE BCA Protein Assay kit (23227, Thermo Scientific). Equal amounts of protein specimens (50 µg/g/lane) were resolved on 12.5% polyacrylamide gels at 130 V and then transferred to polyvinylidene difluoride membranes (Millipore, Billerica, MA, USA). The membranes were blocked with 2.5% non-fat milk at 37°C for 1 h and then probed with primary antibodies against TLR4 (HTA125, 1 : 1000, eBioscience, CA, USA), NF-κB p65 (ab207297, 1 : 1000, Abcam), p-NF-κB p65 (ab76302, 1 : 1000, Abcam), IL-1β (ab254360, 1 : 1000, Abcam), and GAPDH (ab181603, 1 : 10000, Abcam) in a 4°C refrigerator overnight with slow shaking. After rinsing with TBST three times, the membranes were incubated with secondary antibodies (ab288151, 1 : 10000, Abcam) at 37°C for 1 h. After washing the membranes again, the proteins were visualized using the ECL detection system (JMCDVISSP, Millipore) in accordance with the product manuals and quantified with ImageJ software v.1.52v (National Institutes of Health).

Jin Z., Huang Q., Peng J., Liu Z., Hu R., Wu J, & Wang F. (2023). MiR-125a-3p alleviates hyperproliferation of keratinocytes and psoriasis-like inflammation by targeting TLR4/NF-κB pathway. Advances in Dermatology and Allergology/Postȩpy Dermatologii i Alergologii, 40(3), 447-461.

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Histone H4 Regulation of Inflammatory Pathways

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Histone H4 was purchased from Millipore (Billerica, MA, USA). Blocking antibodies against TLR1 (GD2.F4), TLR2 (TL2.1), and TLR4 (HTA125) were purchased from eBioscience (San Diego, CA, USA). Antibodies for TNF-α, IL-1β, P-selectin, Ly6G, H4, Ac-H4(acetyl K5 + K8 + K12 + K16) and blocking antibody against TLR6 (TLR6.127) were purchased from Abcam (Cambridge, UK). An ELISA kit for von Willebrand factor (vWF) and a myeloperoxidase (MPO) detection kit were purchased from Jiancheng Biotech (Nanjing, China). The blocking antibody against Histone H4 (anti-H4) was prepared following the previously described protocol involving autoimmune mice [14 (link)].

Zhang Y., Zhao J., Guan L., Mao L., Li S, & Zhao J. (2020). Histone H4 aggravates inflammatory injury through TLR4 in chlorine gas-induced acute respiratory distress syndrome. Journal of Occupational Medicine and Toxicology (London, England), 15, 31.

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Histone H4 Modulates Immune Response

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Histone H4 was purchased from Millipore (Billerica, MA, USA). Blocking antibodies against TLR1 (GD2.F4), TLR2 (TL2.1), and TLR4 (HTA125) were purchased from eBioscience (San Diego, CA, USA).
Antibodies for TNF-α, IL-1β, P-selectin, Ly6G, H4, Ac-H4(acetyl K5+K8+K12+K16) and blocking antibody against TLR6 (TLR6.127) were purchased from Abcam (Cambridge, UK). An ELISA kit for von Willebrand factor (vWF) and a myeloperoxidase (MPO) detection kit were purchased from Jiancheng Biotech (Nanjing, China). The blocking antibody against Histone H4 (anti-H4) was prepared following the previously described protocol involving autoimmune mice [14] .

Zhang Y., Zhao J., Guan L., Mao L., Li S., & Zhao J. (2020). Histone H4 aggravates inflammatory injury through TLR4 in chlorine gas-induced acute respiratory distress syndrome.

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Histone H4 Modulates Immune Response

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Histone H4 was purchased from Millipore (Billerica, MA, USA). Blocking antibodies against TLR1 (GD2.F4), TLR2 (TL2.1), and TLR4 (HTA125) were purchased from eBioscience (San Diego, CA, USA).
Antibodies for TNF-α, IL-1β, P-selectin, Ly6G, H4, Ac-H4(acetyl K5+K8+K12+K16) and blocking antibody against TLR6 (TLR6.127) were purchased from Abcam (Cambridge, UK). An ELISA kit for von Willebrand factor (vWF) and a myeloperoxidase (MPO) detection kit were purchased from Jiancheng Biotech (Nanjing, China). The blocking antibody against Histone H4 (anti-H4) was prepared following the previously described protocol involving autoimmune mice [14] .

Zhang Y., Zhao J., Guan L., Mao L., Li S., & Zhao J. (2020). Histone H4 aggravates inflammatory injury through TLR4 in chlorine gas-induced acute respiratory distress syndrome.

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Preparation of Monomeric Amyloid-β Peptide

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Aβ1-42 peptide (EZBiolab), Alexa Fluor 488 conjugate of isolectin B 4 from Griffonia simplicifolia (Invitrogen), carboxylate-modified fluorescent microspheres (Invitrogen), cyclo(RGDfV) and cyclo(RADfV) peptides (Bachem), apocynin, Gö6976 and Gö6983 (Calbiochem), TNFα Quantikine Elisa Kit (R&D Systems), function blocking antibodies to TLR4 (HTA-125), TLR2 (T2.5) and isotype control antibodies IgG1, IgG2a (eBioscience). Cell culture reagents were from PAA. Other reagents were from Sigma.
Monomeric amyloid β peptide 1-42 was prepared as described previously (Demuro et al., 2005 , Cizas et al., 2010) : 1.0 mg of lyophilised peptide was dissolved in 400 μL 1,1,1,3,3,3hexafluorisopropanol and solution was incubated for 30-60 minutes at room temperature to ensure complete removal of any pre-existing secondary structure. To prepare monomeric Aβ 1-42 HFIP was removed by evaporation and the peptide was resuspended in DMSO at a concentration of 5 mM.

Neniskyte U., Fricker M, & Brown G.C. (2016). Amyloid β induces microglia to phagocytose neurons via activation of protein kinase Cs and NADPH oxidase. The international journal of biochemistry & cell biology, 81(Pt B).

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