Ab3578

Manufactured by Abcam

Sourced in United Kingdom

Ab3578 is a primary antibody produced in rabbit. It is designed to detect the target protein in various applications.

Automatically generated - may contain errors

Lab products found in correlation

Gelshift chemiluminescent emsa kit, by Active Motif (2 mentions) Ab2774, by Abcam (2 mentions) Ab8901, by Abcam (1 mentions) Midi fl, by 3DHISTECH (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Ab108319, by Abcam (1 mentions) Enhanced chemiluminescence system, by Applygen (1 mentions) Ab196495, by Abcam (1 mentions) Histone h3, by Merck Group (1 mentions) Ab179467, by Abcam (1 mentions)

5 protocols using ab3578

Quantitative Protein-DNA Binding Assay

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EMSA probes were synthesized by Integrated DNA Technologies (IDT). The Gelshift
Chemiluminescent EMSA kit (Active Motif, Carlsbad, CA) was used according to the
manufacturer’s protocols. 3.5 µg of baculovirus recombinant human GR
protein (ab3582) for each condition were incubated with probes (20 fmol) (Supplementary Table 3) for 20 min on
ice. For supershift, anti-GR (ab3578) from Abcam (Cambridge, MA) was incubated for 30 min
after adding probes. The 5% TBE gel was run at 100V for 1 h and then transferred
to nylon membrane. After incubation with streptavidin-HRP conjugate, the membrane was
exposed to film and developed.

Chen Z., Lan X., Wu D., Sunkel B., Ye Z., Huang J., Liu Z., Clinton S.K., Jin V.X, & Wang Q. (2015). Ligand-dependent genomic function of glucocorticoid receptor in triple-negative breast cancer. Nature Communications, 6, 8323.

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Dual Immunofluorescence Staining of Brain Slices

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Brain slices were double-stained for immunohistochemical evaluation using fate-specific antibodies including CRH/BDNF and GR/MR. Briefly, the brain sections were incubated in 3% hydrogen peroxide for 0.5 h and then blocked with normal bovine serum for 0.5 h. The brain sections were washed three times and incubated with rabbit polyclonal anti-CRH (1:1000, ab8901, Abcam, UK), mouse monoclonal anti-BDNF (1:500, ab108319, Abcam, UK), rabbit polyclonal anti-GR (1/200 dilution; ab3578, Abcam, UK), and mouse monoclonal anti-MR (1/200 dilution; ab2774, Abcam, UK) overnight at 4 °C. The slices were then washed with PBS and incubated with Alexa Fluor 647-conjugated donkey anti-rabbit IgG and Alexa Fluor 488-conjugated goat anti-mouse IgG at room temperature for 2 h. Finally, the samples were counterstained with DAPI (Sigma-Aldrich, St. Louis, MO) for 10 min. Digital images of the whole brain sections were obtained by a MIDI FL (3D Histech, Hungary) system.

Zhang B., Bai M., Xu X., Yang M., Niu F., Gao F, & Liu B. (2020). Corticosteroid receptor rebalancing alleviates critical illness-related corticosteroid insufficiency after traumatic brain injury by promoting paraventricular nuclear cell survival via Akt/CREB/BDNF signaling. Journal of Neuroinflammation, 17, 318.

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Gel Shift Assay for GR-DNA Binding

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EMSA probes were synthesized by Integrated DNA Technologies. The Gelshift Chemiluminescent EMSA kit (Active Motif, Carlsbad, CA) was used according to the manufacturer's protocols. Baculovirus recombinant human GR protein (ab3582; 3.5 μg) for each condition were incubated with probes (20 fmol) (Supplementary Table 3) for 20 min on ice. For supershift, anti-GR (ab3578) from Abcam (Cambridge, MA) was incubated for 30 min after adding probes. The 5% TBE gel was run at 100 V for 1 h and then transferred to nylon membrane. After incubation with streptavidin–horseradish peroxidase conjugate, the membrane was exposed to film and developed.

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Immunoblotting Analysis of Ovarian Proteins

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Immunoblotting was performed as described previously (Wu et al. 2014) (link). RIPA lysis buffer was used to extract total protein from mice ovaries. In total, 40 μg of protein were separated in 10% Bis-Tris gels and then transferred onto PVDF membranes. The membranes were blocked for 1 h at room temperature (RT) with TBST (50 mM Tris-HCl, 150 mM NaCl and 0.1% (v/v) Tween-20) containing 5% (w/v) nonfat dried milk and then incubated with rabbit antimouse CYP11A1 (1:600, Proteintech Group, Chicago, IL, USA), rabbit antimouse StAR (1:300, Proteintech Group), rabbit anti-glucocorticoid receptor (1:1000, ab3578, Abcam) and mouse antimouse β-actin (1:2000, Zhongshan, Beijing, China ) antibodies overnight at 4°C. Membranes were then washed with TBST and incubated with HRP-labeled secondary antibody (1:10,000) for 1 h at room temperature. Protein bands on the membrane were visualized by an enhanced chemiluminescence system (Applygen Technologies Inc., Beijing, China) on X-ray film.

Wang S.F., Chen X.H., He B., Yin D.D., Gao H.J., Zhao H.Q., Nan N., Guo S.G., Liu J.B., Wu B, & Xu X.B. (2019). Acute restraint stress triggers progesterone withdrawal and endometrial breakdown and shedding through corticosterone stimulation in mouse menstrual-like model. Reproduction (Cambridge, England), 157(2).

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Protein Expression Profiling in Rat Hippocampus after CCI

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The rats (n=16 per group) were sacri ced by decapitation on day 3 after CCI and the right hippocampus was removed at 4°C. Total (n=8 per group) and nuclear protein (n=8 per group) extraction were performed as previously described [25] . Equal amounts of proteins (30 μg) were separated by SDS-PAGE and then transferred onto polyvinylidene uoride membranes. Blots were incubated in 5% nonfat milk or BSA for 2 hr and then reacted overnight at 4°C with primary antibody P-Akt (ser 473, 1:5000; ab81283), P-Ert (1:1000, CST, #9010), P-CREB (ser133, 1:5000; ab32096), P53 (1:1000, ab131442), Bax (1:1000, ab32503), P-Bad (S136, 1:1000, ab28824), cleaved caspase-3 (1:1000, #9664, CST), Bcl-2 (1:1000; ab196495), MR (1:400, ab2774), GR (1/200 dilution; ab3578), β-Actin (1:5000; ab179467, Abcam, UK), and Histone H3 (1:400; Millipore Co.), followed by incubation with secondary antibodies for 2 hr at room temperature. Blots were visualized using chemiluminescence (Bio Spectrum 500 Imaging System; UVP Co., Upland, CA, USA) and ImageJ software determined the intensity of each band. The percent expression compared with that of the sham controls was calculated for each sample.

Zhang B., Yang M., Yan Q., Xu X., Niu F., Dong J., Zhuang Y., Lu S., Ge Q., & Liu B. (2021). The Dose-dependent Dual Effects of Corticosterone on Spatial Memory Ability After Traumatic Brain Injury Are Mediated by Two Corticosteroid Receptor Systems.

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