Samples of collected tissues and cultured cells were prepared into a homogenate and centrifuged to collect the protein supernatant. Then, after quantifying protein concentrations by using a BCA assay kit (Pierce Biotechnology, Rockford, IL), an appropriate amount of sample protein was resolved by 10% denaturing SDS-PAGE and transferred onto PVDF membranes (Millipore, Bedford, MA), which were then blocked at room temperature for 1 h by using an Odyssey blocking reagent (Li-Cor, Lincoln, NE), incubated overnight in a 4 °C fridge with anti-SIN3A primary antibodies (ab129087, 1:1000 dilution, Abcam, Cambridge, MA), anti-Arc primary antibodies (ab18950, 1:1000 dilution, Abcam, Cambridge, MA), anti-Egr1 primary antibodies (ab133695, 1:1000 dilution, Abcam, Cambridge, MA), anti-Homer1 primary antibodies (ab184955, 1:1000 dilution, Abcam, Cambridge, MA) and anti-Narp primary antibodies (ab191563, 1:1000 dilution, Abcam, Cambridge, MA) and further incubated with HRP-conjugated secondary antibodies (ab6721, 1:2000 dilution, Abcam, Cambridge, MA). The relative protein expression was analyzed using an Odyssey imaging system (Li-Cor, Lincoln, NE).
Ab129087
Manufactured by Abcam
Ab129087 is a lab equipment product manufactured by Abcam. The core function of this product is to [Product description goes here].
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (4 mentions)
Odyssey imaging system, by LI COR (4 mentions)
Odyssey blocking reagent, by LI COR (4 mentions)
Ab6721, by Abcam (4 mentions)
Ab18950, by Abcam (4 mentions)
Ab133695, by Abcam (4 mentions)
Ab184955, by Abcam (4 mentions)
Ab191563, by Abcam (4 mentions)
Bca assay kit, by Thermo Fisher Scientific (4 mentions)
Biotin labeled secondary antibody, by Abcam (4 mentions)
β actin, by Santa Cruz Biotechnology (2 mentions)
Ab13604, by Abcam (1 mentions)
Ab10483, by Abcam (1 mentions)
Sc7872, by Santa Cruz Biotechnology (1 mentions)
Ab8106, by Abcam (1 mentions)
Hrp conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions)
H3k18ac, by Merck Group (1 mentions)
Ab17721, by Abcam (1 mentions)
Ab8580, by Merck Group (1 mentions)
Ab8895, by Abcam (1 mentions)
10 protocols using ab129087
1
Western Blot Analysis of Synaptic Proteins
2
Immunohistochemical Analysis of SIN3A in Rat Hippocampus
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Samples of collected rat hippocampus tissues were sliced into 4 μm sections, fixed using PBS containing 4% paraformaldehyde, and quenched with 3% H2O2 to remove the activity of endogenous peroxidase. Then, after being blocked for 1 h in 5% BSA, the sections were treated overnight in a 4 °C fridge with anti-SIN3A primary antibodies (ab129087, 1:100 dilution, Abcam, Cambridge, MA). After washing and further incubation with biotin-labeled secondary antibodies (1:1000 dilution, Abcam, Cambridge, MA), the slides were incubated with horseradish peroxidase (HRP)-tagged avidin/streptavidin and counter-stained with 4′,6-diamidino-2-phenylindole (DAPI) before the positive expression of SIN3A was analyzed underneath a microscope.
3
Antibody Panel for Epigenetic Regulators
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The list of antibodies we used was as follows: anti-SETDB1 (Upstate 07-378 for NT and Abcam ab12317 for CT), -DICER1 (sc-30226, Santa Cruz), -AGO2 (2897, Cell signaling), -AR (3202, Cell signaling), -PR (3176, Cell signaling), -β-actin (sc-47778, Santa Cruz), -KAP1 (ab10483, Abcam), -EZH2 (3147, Cell signaling), -SIN3A (ab129087, Abcam), -HDAC1 (sc-7872, Santa Cruz), -HDAC2 (sc-7899, Santa Cruz), -MTA2 (ab8106, Abcam), -DNMT3A (D23G1, Cell signaling) and -DNMT3B (ab13604, Abcam).
4
Histone Modification Analysis by Western Blot
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Whole-cell lysate was prepared in 3D-RIPA buffer. Protein (20 µg) was separated by 12% SDS–PAGE and transferred to a polyvinylidene difluoride membrane. Membranes were blocked using 5% nonfat dry milk in PBS containing 0.05% Tween 20, and then incubated with primary antibody overnight at 4 °C. HRP-conjugated secondary antibody was used and detected using the Pierce ECL Western Blot Substrate. Antibodies were obtained from the following sources: H3K9Ac (Abcam, ab32129, 1:1000), H3K4Me2 (Abcam, ab32356, 1:5000), H3K4Me1 (Abcam, ab8895, 1:5000), H3K4Me3 (Abcam, ab8580, 1:5000), H3K14Ac (Millipore, 07–353, 1:5000), H3K18Ac (Millipore, 07–354, 1:5000), LSD1 (Abcam, ab17721, 1:500), HDAC1 (Abcam, ab19845, 1:1000), total H3 (Cell Signaling Technology, 4499 S, 1:1000 or Abcam, ab1791, 1:10,000), β-actin (Santa Cruz Biotechnology, sc47778, 1:2000), CoREST1 (BD Transduction Laboratories, 612146, 1:500), SIN3A (Abcam, ab129087, 1:1000), HRP-conjugated secondary antibodies (Santa Cruz Biotechnology, 1:5000 for β-actin blots, 1:2000 for all others). Blots shown are representative of at least two independent experiments. Uncropped versions of Western blots are provided in the Supporting Information (Supplementary Figs. 32 and 33 ).
5
Immunohistochemical Analysis of SIN3A in Rat Hippocampus
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Samples of collected rat brain tissues in hippocampal CA1 region were sliced into 4 μm sections, xed using PBS containing 4% paraformaldehyde, and quenched with 3% H 2 O 2 to remove the activity of endogenous peroxidase. Then, after being blocked for 1 h in 5% BSA, the sections were treated overnight in a 4°C fridge with anti-SIN3A primary antibodies (ab129087, 1:100 dilution, Abcam, Cambridge, MA).
After washing and further incubation with biotin-labeled secondary antibodies (1:1000 dilution, Abcam, Cambridge, MA), the slides were incubated with horseradish peroxidase (HRP)-tagged avidin/streptavidin and counter-stained with 4',6-diamidino-2-phenylindole (DAPI) before the positive expression of SIN3A was analyzed underneath a microscope.
After washing and further incubation with biotin-labeled secondary antibodies (1:1000 dilution, Abcam, Cambridge, MA), the slides were incubated with horseradish peroxidase (HRP)-tagged avidin/streptavidin and counter-stained with 4',6-diamidino-2-phenylindole (DAPI) before the positive expression of SIN3A was analyzed underneath a microscope.
6
Immunohistochemical Analysis of SIN3A
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Samples of collected rat hippocampus tissues were sliced into 4 μm sections, xed using PBS containing 4% paraformaldehyde, and quenched with 3% H 2 O 2 to remove the activity of endogenous peroxidase.
Then, after being blocked for 1 h in 5% BSA, the sections were treated overnight in a 4°C fridge with anti-SIN3A primary antibodies (ab129087, 1:100 dilution, Abcam, Cambridge, MA). After washing and further incubation with biotin-labeled secondary antibodies (1:1000 dilution, Abcam, Cambridge, MA), the slides were incubated with horseradish peroxidase (HRP)-tagged avidin/streptavidin and counter-stained with 4',6-diamidino-2-phenylindole (DAPI) before the positive expression of SIN3A was analyzed underneath a microscope.
Then, after being blocked for 1 h in 5% BSA, the sections were treated overnight in a 4°C fridge with anti-SIN3A primary antibodies (ab129087, 1:100 dilution, Abcam, Cambridge, MA). After washing and further incubation with biotin-labeled secondary antibodies (1:1000 dilution, Abcam, Cambridge, MA), the slides were incubated with horseradish peroxidase (HRP)-tagged avidin/streptavidin and counter-stained with 4',6-diamidino-2-phenylindole (DAPI) before the positive expression of SIN3A was analyzed underneath a microscope.
7
Western Blot Analysis of Neuronal Proteins
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Samples of collected tissues and cultured cells were prepared into a homogenate and centrifuged to collect the protein supernatant. Then, after quantifying protein concentrations by using a BCA assay kit (Pierce Biotechnology, Rockford, IL), an appropriate amount of sample protein was resolved by 10% denaturing SDS-PAGE and transferred onto PVDF membranes (Millipore, Bedford, MA), which were then blocked at room temperature for 1 h by using an Odyssey blocking reagent (Li-Cor, Lincoln, NE), incubated overnight in a 4 o C fridge with anti-SIN3A primary antibodies (ab129087, 1:1000 dilution, Abcam, Cambridge, MA), anti-Arc primary antibodies (ab18950, 1:1000 dilution, Abcam, Cambridge, MA), anti-Egr1 primary antibodies (ab133695, 1:1000 dilution, Abcam, Cambridge, MA), anti-Homer1 primary antibodies (ab184955, 1:1000 dilution, Abcam, Cambridge, MA) and anti-Narp primary antibodies (ab191563, 1:1000 dilution, Abcam, Cambridge, MA) and further incubated with HRP-conjugated secondary antibodies (ab6721, 1:2000 dilution, Abcam, Cambridge, MA). The relative protein expression was analyzed using an Odyssey imaging system (Li-Cor, Lincoln, NE).
8
Western Blot Analysis of Neural Proteins
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Samples of collected tissues and cultured cells were prepared into a homogenate and centrifuged to collect the protein supernatant. Then, after quantifying protein concentrations by using a BCA assay kit (Pierce Biotechnology, Rockford, IL), an appropriate amount of sample protein was resolved by 10% denaturing SDS-PAGE and transferred onto PVDF membranes (Millipore, Bedford, MA), which were then blocked at room temperature for 1 h by using an Odyssey blocking reagent (Li-Cor, Lincoln, NE), incubated overnight in a 4 o C fridge with anti-SIN3A primary antibodies (ab129087, 1:1000 dilution, Abcam, Cambridge, MA), anti-Arc primary antibodies (ab18950, 1:1000 dilution, Abcam, Cambridge, MA), anti-Egr1 primary antibodies (ab133695, 1:1000 dilution, Abcam, Cambridge, MA), anti-Homer1 primary antibodies (ab184955, 1:1000 dilution, Abcam, Cambridge, MA) and anti-Narp primary antibodies (ab191563, 1:1000 dilution, Abcam, Cambridge, MA) and further incubation with HRP-conjugated secondary antibodies (ab6721, 1:2000 dilution, Abcam, Cambridge, MA). The relative protein expression was analyzed by using an Odyssey imaging system (Li-Cor, Lincoln, NE).
9
Immunohistochemical Analysis of SIN3A
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples of collected rat hippocampus tissues were sliced into 4 μm sections, xed using PBS containing 4% paraformaldehyde, and quenched with 3% H 2 O 2 to remove the activity of endogenous peroxidase.
Then, after being blocked for 1 h in 5% BSA, the sections were treated overnight in a 4°C fridge with anti-SIN3A primary antibodies (ab129087, 1:100 dilution, Abcam, Cambridge, MA). After washing and further incubation with biotin-labeled secondary antibodies (1:1000 dilution, Abcam, Cambridge, MA), the slides were incubated with horseradish peroxidase (HRP)-tagged avidin/streptavidin and counter-stained with 4',6-diamidino-2-phenylindole (DAPI) before the positive expression of SIN3A was analyzed underneath a microscope.
Then, after being blocked for 1 h in 5% BSA, the sections were treated overnight in a 4°C fridge with anti-SIN3A primary antibodies (ab129087, 1:100 dilution, Abcam, Cambridge, MA). After washing and further incubation with biotin-labeled secondary antibodies (1:1000 dilution, Abcam, Cambridge, MA), the slides were incubated with horseradish peroxidase (HRP)-tagged avidin/streptavidin and counter-stained with 4',6-diamidino-2-phenylindole (DAPI) before the positive expression of SIN3A was analyzed underneath a microscope.
10
Western Blot Analysis of Neuronal Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples of collected tissues and cultured cells were prepared into a homogenate and centrifuged to collect the protein supernatant. Then, after quantifying protein concentrations by using a BCA assay kit (Pierce Biotechnology, Rockford, IL), an appropriate amount of sample protein was resolved by 10% denaturing SDS-PAGE and transferred onto PVDF membranes (Millipore, Bedford, MA), which were then blocked at room temperature for 1 h by using an Odyssey blocking reagent (Li-Cor, Lincoln, NE), incubated overnight in a 4 o C fridge with anti-SIN3A primary antibodies (ab129087, 1:1000 dilution, Abcam, Cambridge, MA), anti-Arc primary antibodies (ab18950, 1:1000 dilution, Abcam, Cambridge, MA), anti-Egr1 primary antibodies (ab133695, 1:1000 dilution, Abcam, Cambridge, MA), anti-Homer1 primary antibodies (ab184955, 1:1000 dilution, Abcam, Cambridge, MA) and anti-Narp primary antibodies (ab191563, 1:1000 dilution, Abcam, Cambridge, MA) and further incubated with HRP-conjugated secondary antibodies (ab6721, 1:2000 dilution, Abcam, Cambridge, MA). The relative protein expression was analyzed using an Odyssey imaging system (Li-Cor, Lincoln, NE).
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