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Gtx109242

Manufactured by GeneTex
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Sourced in United States

The GTX109242 is a high-performance laboratory centrifuge designed for a variety of applications. It features a compact and durable construction, providing reliable and consistent operation. The centrifuge can accommodate a range of sample volumes and tube sizes, making it a versatile tool for various laboratory tasks.

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Lab products found in correlation

Ab15323, by Abcam (3 mentions) Alexa fluor 568 goat anti mouse igg, by Thermo Fisher Scientific (2 mentions) Goat anti rabbit igg fitc, by Jackson ImmunoResearch (2 mentions) Ab129372, by Abcam (2 mentions) Alexa 647, by Thermo Fisher Scientific (2 mentions) D3571, by Thermo Fisher Scientific (2 mentions) H 3300, by Vector Laboratories (2 mentions) α actin fitc acta2, by Merck Group (2 mentions) Irak phosphor t209, by Abcam (2 mentions) Ab15580, by Abcam (2 mentions) Ab192256, by Abcam (2 mentions) Prolong gold antifade, by Thermo Fisher Scientific (2 mentions) Ab9661, by Cell Signaling Technology (2 mentions) Cl8942ap, by Abcam (2 mentions) Cf488a, by Biotium (2 mentions) Ab6673, by Abcam (2 mentions) Pecam 1, by Santa Cruz Biotechnology (2 mentions) Sc 8409, by Santa Cruz Biotechnology (1 mentions) Ab181553, by Abcam (1 mentions) Yt4689, by Immunoway (1 mentions)

7 protocols using gtx109242

1

Protein Expression in Ischemic Penumbra

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Proteins from ischemic penumbral tissues and HAPI cells were used for Western blot analysis. SDS-PAGE was performed, and the resolved proteins were transferred onto polyvinylidene fluoride (PVDF) membranes by electroblotting. The membranes were blocked at Tris–buffered saline (TBST) containing 5% non-fat milk powder for 2 h and then incubated with the following primary antibodies overnight at 4°C: Arg1 (1:500, GTX109242, GeneTex), CD163 (1:500, WH112776, ABclonal), iNOS (1:500, GTX130246, GeneTex), CD86 (1:500, WH141312, ABclonal), P62 (1:200, WH146703, ABclonal), IRF5 (1:1000, ab181553, Abcam), TRAF6 (1:200, sc-8409, Santa Cruz), IKKαβ (1:200, BM4499, Boster), IL-10 (1:200, 20850-1-AP, Proteintech), IL-4 (1:500, 66142-1-lg, Proteintech), TNF-α (1:200, YT4689, ImmunoWay), and IL-1β (1:500, DF6251, Affinity). After being washed with TBST 3 times, the membranes were incubated with horseradish peroxidase (HRP)-conjugated goat anti-rabbit/mouse secondary antibodies for 2 h at RT. β-actin was used as a loading control. Immunoreactive bands were detected by an enhanced chemiluminescence (ECL) detection system. The gray values of the protein bands were quantified by Image Lab.
Wang T., Zhao N., Peng L., Li Y., Huang X., Zhu J., Chen Y., Yu S, & Zhao Y. (2020). DJ-1 Regulates Microglial Polarization Through P62-Mediated TRAF6/IRF5 Signaling in Cerebral Ischemia-Reperfusion. Frontiers in Cell and Developmental Biology, 8, 593890.
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2

Quantifying Macrophage Polarization Markers

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Macrophages seeded on tissue arrays in M1 or M2 polarizing cytokines as described above were evaluated for expression of the M2 and M1 markers Arginase-1 and iNOS. After 24 hours, macrophages were fixed for 10 min in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100 in TBS for 10 min. Nonspecific protein interactions were blocked with 1% bovine serum albumin, 2% horse serum in 0.05% Tween-20. Tissue microarrays were then incubated with primary antibodies against Arginase-1 (rabbit polyclonal diluted 1:100, GTX109242, GeneTex) and iNOS (mouse monoclonal [4E5] diluted 1:300, ab129372, abcam) diluted in 1% bovine serum albumin in TBS overnight at 4°C. Arrays were washed and probed with FITC goat anti-rabbit IgG (1:250, Jackson Immuno) and Alexa Fluor-568 goat anti-mouse IgG (1:250, Invitrogen) secondary antibodies diluted in 1% bovine serum albumin for 1 hour. ECM autofluorescence was then blocked by treatment with 0.1% Sudan Black B in 70% ethanol for 20 min. Arrays were washed, coverslipped, and imaged. Variations in background autofluorescence were corrected by subtracting the mean background intensity of acellular regions of each tissue array spot using ImageJ software.
Beachley V.Z., Wolf M.T., Sadtler K., Manda S.S., Jacobs H., Blatchley M., Bader J.S., Pandey A., Pardoll D, & Elisseeff J.H. (2015). Tissue matrix arrays for high throughput screening and systems analysis of cell function. Nature methods, 12(12), 1197-1204.
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3

Protein Expression Analysis in Cells

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Cells were washed twice with PBS and resuspended in lysis buffer (50 mM Tris–HCl pH 8.0, 1 mM EDTA, 250 mM NaCl, 1% NP-40 and 0.5% Na-Deoxycholate). Cell lysates were then shaken for 30 min on an orbital shaker at 4 °C and centrifuged for 20 min at 12,000×g and the protein containing supernatant was collected. Protein concentration of cell lysates was estimated using a commercial kit (Bio-Rad, USA). SDS-PAGE and Western blot were performed to determine the expression of Arg-1 (GTX109242, Gene Tex, USA), iNOS (GTX31048, Gene Tex, USA), Hif-1α (2015–1, Epitomics, USA), IRAK1 (4504, Cell signaling, USA), IRAK2 (4367, Cell signaling, USA), IRAK4 (4363, Cell signaling, USA), IRAK-M (4369, Cell signaling, USA), pERK1/2 (2219–1, Epitomics, USA), pSTAT1 (7649, Cell signaling, USA), pSTAT6 (9361, Cell signaling, USA), STAT1 (10144–2-AP, Proteintech, USA), STAT6 (51073–1-AP, Proteintech, USA), VEGF (sc-7269, Santa cruz, USA), β-actin (M2010S, Abmart, China).
Shen P., Li Q., Ma J., Tian M., Hong F., Zhai X., Li J., Huang H, & Shi C. (2017). IRAK-M alters the polarity of macrophages to facilitate the survival of Mycobacterium tuberculosis. BMC Microbiology, 17, 185.
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4

Immunohistochemical Analysis of Arg-1 and iNOS

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Slides were dried overnight and fixed in acetone (10 minutes, −20°C), then rehydrated with PBS (10 minutes) and blocked with 0.5% fish gelatin (G‐7765; Sigma‐Aldrich, St. Louis, MO), 1% BSA, and 0.3% Triton X‐100 (blocking buffer, 1 hour, room temperature). Sections were treated with antibodies to Arg‐1 (GTX109242, 1:400 in blocking buffer; GeneTex, Irvine, CA) or iNOS (ab15323, 1:50 in blocking buffer; Abcam) overnight at 4°C. Alexa 488–labeled goat anti‐rabbit (A‐11034; Invitrogen, Carlsbad, CA) was used as the secondary antibody (1:500, 60 minutes, room temperature). Nuclei were visualized with 4′,6‐diamidino‐2‐phenylindole (1:1000, 15 minutes, room temperature).
Harmon E.Y., Fronhofer V., Keller R.S., Feustel P.J., Zhu X., Xu H., Avram D., Jones D.M., Nagarajan S, & Lennartz M.R. (2014). Anti‐Inflammatory Immune Skewing Is Atheroprotective: Apoe−/−FcγRIIb−/− Mice Develop Fibrous Carotid Plaques. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 3(6), e001232.
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5

Multiparametric Immunohistochemistry Analysis

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BCA sections were de-paraffinized and rehydrated in xylene and ethanol series. After antigen retrieval (H-3300, Vector Laboratories), sections were blocked with fish skin gelatin–PBS (6 g/L) containing 10% horse serum for 1 hour at room temperature. Slides were incubated with the following antibodies: mouse monoclonal SM α-actin-FITC (ACTA2) (4.4 μg/mL, clone 1A4, Sigma Aldrich), goat polyclonal anti-GFP (4 μg/mL, ab6673, Abcam) for detection of YFP, LGALS3 (2 μg/mL, Cedarlane CL8942AP), RUNX2 (1.374 μg/mL, ab192256, Abcam), Ki67 (4 μg/mL, ab15580, Abcam), PECAM-1 (1 μg/mL, Santa Cruz), IRAK phosphor-T209 (4 μg/mL, Abcam), iNOS (0.52 μg/mL, ab15323, Abcam), and Arg1 (9.2 μg/mL, GTX109242, GeneTex). The secondary antibodies were donkey anti-rat conjugated to Alexa 555 (5 μg/mL, Abcam), donkey anti-rat conjugated to Alexa 647 (5 μg/mL, Abcam), donkey anti-goat conjugated to Alexa 555 (5 μg/mL, Invitrogen), and donkey anti-goat conjugated to Alexa 647 (4 μg/mL, Invitrogen). Apoptosis was assessed by TUNEL (CF488A, Biotium) and cleaved caspase 3 staining (0.84 ug/mL, ab9661, Cell Signaling Technology). IL-1β antibody was detected using a donkey anti-mouse IgG2a-Alexa 488 antibody. DAPI (0.05 mg/mL, D3571, ThermoFisher Scientific) was used as a nuclear counterstain and slides were mounted using Prolong Gold Antifade (Invitrogen).
Gomez D., Baylis R.A., Durgin B.G., Newman A.A., Alencar G.F., Mahan S., St. Hilaire C., Muller W., Waisman A., Francis S.E., Pinteaux E., Randolph G.J., Gram H, & Owens G.K. (2018). Interleukin-1β promotes atheroprotective changes of advanced atherosclerotic lesions in mice. Nature medicine, 24(9), 1418-1429.
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6

Multiparametric Immunohistochemistry Analysis

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BCA sections were de-paraffinized and rehydrated in xylene and ethanol series. After antigen retrieval (H-3300, Vector Laboratories), sections were blocked with fish skin gelatin–PBS (6 g/L) containing 10% horse serum for 1 hour at room temperature. Slides were incubated with the following antibodies: mouse monoclonal SM α-actin-FITC (ACTA2) (4.4 μg/mL, clone 1A4, Sigma Aldrich), goat polyclonal anti-GFP (4 μg/mL, ab6673, Abcam) for detection of YFP, LGALS3 (2 μg/mL, Cedarlane CL8942AP), RUNX2 (1.374 μg/mL, ab192256, Abcam), Ki67 (4 μg/mL, ab15580, Abcam), PECAM-1 (1 μg/mL, Santa Cruz), IRAK phosphor-T209 (4 μg/mL, Abcam), iNOS (0.52 μg/mL, ab15323, Abcam), and Arg1 (9.2 μg/mL, GTX109242, GeneTex). The secondary antibodies were donkey anti-rat conjugated to Alexa 555 (5 μg/mL, Abcam), donkey anti-rat conjugated to Alexa 647 (5 μg/mL, Abcam), donkey anti-goat conjugated to Alexa 555 (5 μg/mL, Invitrogen), and donkey anti-goat conjugated to Alexa 647 (4 μg/mL, Invitrogen). Apoptosis was assessed by TUNEL (CF488A, Biotium) and cleaved caspase 3 staining (0.84 ug/mL, ab9661, Cell Signaling Technology). IL-1β antibody was detected using a donkey anti-mouse IgG2a-Alexa 488 antibody. DAPI (0.05 mg/mL, D3571, ThermoFisher Scientific) was used as a nuclear counterstain and slides were mounted using Prolong Gold Antifade (Invitrogen).
Gomez D., Baylis R.A., Durgin B.G., Newman A.A., Alencar G.F., Mahan S., St. Hilaire C., Muller W., Waisman A., Francis S.E., Pinteaux E., Randolph G.J., Gram H, & Owens G.K. (2018). Interleukin-1β promotes atheroprotective changes of advanced atherosclerotic lesions in mice. Nature medicine, 24(9), 1418-1429.
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7

Quantifying Macrophage Polarization Markers

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Macrophages seeded on tissue arrays in M1 or M2 polarizing cytokines as described above were evaluated for expression of the M2 and M1 markers Arginase-1 and iNOS. After 24 hours, macrophages were fixed for 10 min in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100 in TBS for 10 min. Nonspecific protein interactions were blocked with 1% bovine serum albumin, 2% horse serum in 0.05% Tween-20. Tissue microarrays were then incubated with primary antibodies against Arginase-1 (rabbit polyclonal diluted 1:100, GTX109242, GeneTex) and iNOS (mouse monoclonal [4E5] diluted 1:300, ab129372, abcam) diluted in 1% bovine serum albumin in TBS overnight at 4°C. Arrays were washed and probed with FITC goat anti-rabbit IgG (1:250, Jackson Immuno) and Alexa Fluor-568 goat anti-mouse IgG (1:250, Invitrogen) secondary antibodies diluted in 1% bovine serum albumin for 1 hour. ECM autofluorescence was then blocked by treatment with 0.1% Sudan Black B in 70% ethanol for 20 min. Arrays were washed, coverslipped, and imaged. Variations in background autofluorescence were corrected by subtracting the mean background intensity of acellular regions of each tissue array spot using ImageJ software.
Beachley V.Z., Wolf M.T., Sadtler K., Manda S.S., Jacobs H., Blatchley M., Bader J.S., Pandey A., Pardoll D, & Elisseeff J.H. (2015). Tissue matrix arrays for high throughput screening and systems analysis of cell function. Nature methods, 12(12), 1197-1204.
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