Ab150130

Manufactured by Abcam

Sourced in United Kingdom

Ab150130 is a primary antibody for detecting the GAPDH protein. It is a rabbit monoclonal antibody that can be used in various immunoassays, including Western blotting and immunohistochemistry. The antibody recognizes the GAPDH protein, which is a key enzyme involved in glycolysis.

Automatically generated - may contain errors

Lab products found in correlation

Fv1000, by Olympus (1 mentions) Ab9957, by Abcam (1 mentions) Af459, by R&D Systems (1 mentions) Ab150073, by Abcam (1 mentions) Sc 53029, by Santa Cruz Biotechnology (1 mentions) Glass bottom dishes, by MatTek (1 mentions) Slowfade antifade, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by MP Biomedicals (1 mentions) Ab150079, by Abcam (1 mentions) Ab87645, by Abcam (1 mentions) Sc28999, by Santa Cruz Biotechnology (1 mentions) Sc30004, by Santa Cruz Biotechnology (1 mentions) M 300, by Santa Cruz Biotechnology (1 mentions) Sc 16240, by Santa Cruz Biotechnology (1 mentions) Ab22758, by Abcam (1 mentions) Ab24011, by Abcam (1 mentions) Ab177487, by Abcam (1 mentions) H 114, by Santa Cruz Biotechnology (1 mentions) Ab150077, by Abcam (1 mentions) Ab150115, by Abcam (1 mentions)

5 protocols using ab150130

Localization of RANKL and OPG in Osteoblasts

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To detect and locate the expression changes of nuclear factor кB receptor activating factor ligand (RANKL) and osteoprotegerin (OPG) under the intervention of Ti CM and SR in osteoblasts, we performed cellular immunofluorescence combined with confocal laser microscopy assay. The cells were grouped and treated as described above. Afterward, the cells were fixed with 4% paraformaldehyde for 10 min and then permeabilized with 0.1% Triton X-100 for 20 min at room temperature. Non-specific binding sites were blocked by normal donkey serum. The cells were incubated with rabbit anti-mouse RANKL (ab9957, Abcam, 1 μg/ml) and goat anti-mouse OPG (AF459, R&D systems, 1 μg/ml) antibody overnight at 4°C, after which they were washed three times with PBS and incubated with donkey anti-rabbit IgG H&L (Alexa Fluor 488, ab150073, Abcam, 1:500) and donkey anti-goat IgG H&L (Alexa Fluor 555, ab150130, Abcam, 1:500) fluorescent secondary antibodies for 30 min at 37°C. Finally, the cells were counterstained with DAPI in the dark and photographed under 40-fold using a confocal laser microscope system (FV-1000, Olympus, Japan).

Wang B., Guo H., Geng T., Sun K., Zhang L., Lu Z, & Jin Q. (2021). The effect of strontium ranelate on titanium particle-induced periprosthetic osteolysis regulated by WNT/β-catenin signaling in vivo and in vitro. Bioscience Reports, 41(1), BSR20203003.

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Immunofluorescence Staining of 14-3-3 and Tubulin

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Cells were allowed to grow on glass-bottom dishes (MatTek) and were washed with PB buffer (10 mM KH₂PO₄/Na₂HPO₄, pH 6.5) before fixation. Cells were fixated by incubation for 15 min with PB containing 4% paraformaldehyde, washed with PB, and permeabilized by two 10-min incubations in PB containing 0.1% Triton X-100. Subsequently cells were blocked for 30 min in PB containing 1% wt/vol bovine serum albumin (BSA) and 0.1% Tween, incubated with 14-3-3 antibody (ab77187 [Abcam, Cambridge, UK], 1:200 diluted in 1% BSA, 0.1% Tween PB) or α-tubulin antibody (sc53029 [Santa Cruz, Heidelberg, Germany], 1:50 diluted in 1%BSA, 0.1% Tween PB) for 1 h, washed three times with PB buffer, and finally incubated for 1 h in the dark with a secondary antibody (ab150130 [Abcam] or sc2782 [SantaCruz], 1:200 dilution with 1% BSA, 0.1% Tween, PB). Staining of DNA was performed with 20 μg/ml DAPI solution in PB for 10 min.

Plak K., Keizer-Gunnink I., van Haastert P.J, & Kortholt A. (2014). Rap1-dependent pathways coordinate cytokinesis in Dictyostelium. Molecular Biology of the Cell, 25(25), 4195-4204.

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Immunofluorescence Staining of Connexin-43

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Slides were deparaffinized in xylene and rehydrated in graded ethanol. After washing with TBS-T (1× TBS, 0.025% Triton), slides were permeabilized in 50% methanol followed by heat induced epitope retrieval by microwaving for 25 min in sodium citrate buffer (10 mM sodium citrate, 0.05% Tween 20, pH 6.0). Slides were washed again with TBS-T and blocked (1× TBS with 2.5% horse serum and 0.2% Triton) for 1 h at RT and incubated at 4 °C overnight with the following primary antibodies: α-Cx43 (1:50, ab87645 Abcam) and α-Cx43 pY313 (1:100). The next day, sections were washed 3 × 10 min with 1× TBS-T, incubated with secondary antibody (ab150130 and ab150079, Abcam) for 1 h at RT, stained with DAPI (100 ng/mL) (MP Biomedicals) for 10 min, and then washed 3 × 10 min with 1× TBS-T. Coverslips were mounted on a drop of SlowFade anti-fade (Life Tech), sealed with clear nail polish, and imaged.

Zheng L., Li H., Cannon A., Trease A.J., Spagnol G., Zheng H., Radio S., Patel K., Batra S, & Sorgen P.L. (2018). Phosphorylation of Cx43 residue Y313 by Src contributes to blocking the interaction with Drebrin and disassembling gap junctions. Journal of molecular and cellular cardiology, 126, 36-49.

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Comprehensive Antibody Immunodetection Protocol

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A goat polyclonal antibody against TLR4 (sc-16,240), a rabbit polyclonal IgG antibody (sc-28,999) against TLR3 (M-300), and a rabbit polyclonal IgG antibody (sc-30,004) against TLR7 (H-114) were purchased from Santa Cruz Biotechnology (TX, USA). A rabbit polyclonal antibody against C23 (ab22758), a mouse monoclonal antibody against F protein (RSV3216 (B016), ab24011), an anti-NeuN antibody (EPR12763, neuronal marker, ab177487), a donkey anti-goat IgG heavy and light chain (H&L) antibody (Alexa Fluor 555, ab150130) against TLR4, a goat anti-rabbit IgG H&L antibody (Alexa Fluor 488, ab150077) against C23, and a goat anti-mouse IgG H&L antibody (Alexa Fluor 647, ab150115) against RSV F were purchased from Abcam (Cambridge, UK). The Annexin V-FITC apoptosis kit (BB-4101) was obtained from BestBio (Shanghai, China), and enzyme-linked immunosorbent assay (ELISA) kits for IL-6 and TNF-α were obtained from Hermes Criterion Biotechnology (Vancouver, Canada).

Yuan X., Hu T., He H., Qiu H., Wu X., Chen J., Wang M., Chen C, & Huang S. (2018). Respiratory syncytial virus prolifically infects N2a neuronal cells, leading to TLR4 and nucleolin protein modulations and RSV F protein co-localization with TLR4 and nucleolin. Journal of Biomedical Science, 25, 13.

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Immunostaining of Muscle Tissue for Macrophages and Interleukin-6

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Frozen tissues of 6 μm thickness were obtained from gastrocnemius. Frozen tissues were obtained by soaking samples into isopentane which precooled at −150 °C. Serial sections were fixed in 4% paraformaldehyde for 7 min and blocking by 1% bovine serum albumin (BSA) solution for 30 min at room temperature. Anti-F4/80 (ab6640; Abcam, Cambridge, U.K.) and anti-IL-6 (AF406; R&D Systems, Minneapolis, MN, USA) primary antibodies diluted in 1% BSA solution were incubated with the sections overnight at 4 °C. Alexa Fluor 488 donkey anti-rat IgG (A-21208; Thermo Fisher Scientific, Rockford, IL, USA) and Alexa Fluor 555 donkey anti-goat IgG (ab150130; Abcam, Cambridge, U.K.) antibodies diluted in 1% BSA solution were incubated with the sections for 1 h at room temperature. The concentration of the antibodies was 15 μg/mL for IL-6 and 10 μg/mL for F4/80, Alexa Fluor 488 and Alexa Fluor 555.
The stained sections of the muscle tissue were visualized by fluorescence microscopy (KEYENCE, Osaka, Japan). F4/80-positive and F4/80 and IL-6 double-positive cells were counted in three random 200× magnification fields per slide to derive the average value for each section. F4/80-positive cells and F4/80 and IL-6 double-positive cells were detected with visual judgment of the observer.

Tominaga T., Ma S., Saitou K, & Suzuki K. (2019). Glucose Ingestion Inhibits Endurance Exercise-Induced IL-6 Producing Macrophage Infiltration in Mice Muscle. Nutrients, 11(7), 1496.

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