Ab14939

Manufactured by Abcam

Sourced in United States

Ab14939 is a primary antibody product. The core function of this product is to serve as a research tool for the detection and analysis of target proteins in various biological samples.

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Lab products found in correlation

5 protocols using ab14939

Immunohistochemical Analysis of CSN3 and SOCS3

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Mice were anesthetized and intracardially perfused with 0.9% sodium chloride followed by 4% paraformaldehyde. Brains were removed and post-fixed in 4% PFA overnight at 4°C. Coronal brain sections (10 μm) were cut on a freezing microtome. The sections were stained overnight at 4°C using anti-CSN3 antibody (1:50, ab229807, Abcam) or anti-SOCS3 (1:50, ab14939, Abcam). Then, the sections were incubated using an appropriate secondary antibody for 1 h at room temperature, and the sections were imaged by microscope.

Liang E., Li X., Fu W., Zhao C., Yang B, & Yang Z. (2021). COP9 Signalosome Subunit 3 Restricts Neuroinflammatory Responses During Cerebral Ischemia/Reperfusion Injury Through Stabilizing Suppressor of Cytokine Signaling 3 Protein. Neuropsychiatric Disease and Treatment, 17, 1217-1227.

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Immunocytochemistry Assay for SOCS3 in BV2 Cells

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After OGD/R-0h/24h treatment, BV2 cells were fixed with 4% paraformaldehyde and then permeabilized with Triton X-100. Subsequently, cells were incubated with anti-SOCS3 antibody (ab14939, Abcam) at 4°C overnight, and then incubated with secondary antibodies [goat anti-mouse Alexa Fluor 594 (A11032, Invitrogen)] at room temperature for 1 h. Nuclei were stained with DAPI (Roche, USA) for 10 min at room temperature. The cells were examined with a ﬂuorescence microscope (Nikon TE300, Japan).

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Investigating HCMV-induced SOCS3 expression

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NPCs were seeded on poly-D-lysine-coated coverslips in 12-well plates and infected with HCMV after attachment. The coverslips were harvested and fixed with 4% paraformaldehyde at the indicated times post-infection. PBS-perfused Brains were fixed in 4% paraformaldehyde and prepared for IFA. anti-SOCS3 (IgG2b; ab14939, Abcam), anti-SOCS3 (Rabbit; ab16030, Abcam), anti-HA (Rabbit; Covance) and anti-IE1/2 (IgG1; Virusys) were detected by primary antibodies and appropriate secondary antibodies [35 (link),64 (link)]. The secondary antibodies included TRITC-mouse IgG2b (1090–03, Southern Biotech), Alexa Fluor 488-mouse IgG1 (A-21121, Invitrogen), Alexa Fluor 647-rabbit (A-21247, Invitrogen), and Nuclei were counterstained with DAPI (4′,6-diamidino-2- phenylindole) (D9542, Merck).

Wang X.Z., Wen L., Zhou Y.P., Huang S.N., Yang B., Cheng S., Zeng W.B., Mei M.J., Sun J.Y., Jiang X., Cheng H, & Luo M.H. (2023). Human cytomegalovirus pUL97 upregulates SOCS3 expression via transcription factor RFX7 in neural progenitor cells. PLOS Pathogens, 19(2), e1011166.

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Western Blot Analysis of SAH in Rats

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Western blot was performed as previously reported.^{19 (link)} Briefly, the rats were sacrificed at 3h, 6h, 12h, 24h, and 72h after SAH. Rats were anesthetized and perfused with 0.1M PBS, and the left hemispheres were immediately harvested. Samples were mixed with RIPA, after centrifuged at 14000g for 20min (4°C), the supernatants were collected. A protein assay kit was used for measuring the protein concentration of the samples. Equal amounts of proteins (40μg) were loaded onto the SDS-PAGE gel and transferred onto polyvinylidene fluoride membranes. Membranes were incubated with primary antibodies against EZH2 (1;1000, ab191080, Abcam, USA), H3K27Me3 (1:1000,ab6002, Abcam, USA), H3 (1:1000, ab1791, Abcam, USA), SOCS3(1:1000, ab14939, Abcam, USA), TRAF6 (1:1000, ab227560, Abcam, USA), NF-κB(1:1000, ab16502, Abcam, USA), IL-1β (1:1000, ab9722, Abcam, USA), IL-6 (1:1000, ab6672, Abcam, USA), TNF-α (1:1000, ab1793, Abcam, USA), and IL-10 (1:1000, ab33471, Abcam, USA) overnight at 4°C. On the following day, secondary antibodies were incubated at room temperature for 2h. The bands were visualized by ECL reagents. The proteins band densities were analyzed with Image J software.

Luo Y., Fang Y., Kang R., Lenahan C., Gamdzyk M., Zhang Z., Okada T., Tang J., Chen S, & Zhang J.H. (2020). Inhibition of EZH2 attenuates neuroinflammation via H3k27me3/SOCS3/TRAF6/NF-κB in a rat model of subarachnoid hemorrhage. Stroke, 51(11), 3320-3331.

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Western Blot Analysis of SOCS3 and STAT3

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After treatments, BV2 cells were collected and lysed using NP40 lysis buffer (Beyotime Biotechnology, China) on ice. Lysates were quantified by the BCA kit (Beyotime Biotechnology, China), separated by 10% SDS-PAGE gel and then transferred onto polyvinylidene difluoride membranes. Membranes were blocked, incubated sequentially with primary and secondary antibodies. The immunoblots were detected using chemiluminescence (ECL Plus detection system). Bands were quantified using Image J software. The primary antibodies used were as follows: SOCS3 (1:1000, ab14939, Abcam), STAT3 (1:1000, ab68153, Abcam), phosphorylated STAT3 (pSTAT3, 1:1000, ab76315, Abcam), flag (1:1000, 80010-1-RR, ProteinTech), GAPDH (1:1000, 60004-1-Ig, ProteinTech), Myc (1:1000, 16286-1-AP, ProteinTech), CSN2 (1:1000, 10969-2-AP, ProteinTech), CSN3 (1:1000, ab229807, Abcam), CSN4 (1:1000, 10464-1-AP, ProteinTech), CSN5 (1:1000, 27511-1-AP, ProteinTech), CSN6 (1:1000, ab77299, Abcam), CSN7 (1:1000, ab133548, Abcam), CSN8 (1:1000, 10089-2-AP, ProteinTech). The secondary antibodies used in our work were as follows: goat anti-rabbit IgG-HRP (1:3000, SA00001-2, ProteinTech), goat anti-mouse IgG-HRP (1:3000, SA00001-1, ProteinTech).

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