rabbit polyclonal anti hdac2  (Proteintech)

Journal: Canadian Respiratory Journal

Article Title: Role of Erythromycin-Regulated Histone Deacetylase-2 in Benign Tracheal Stenosis

doi: 10.1155/2020/4213807

Figure Lengend Snippet: The expression of HDAC2 in the different groups determined by immunofluorescence. Image scale bar, 100 µ m. Red fluorescence (Cy3 staining) for the detection of the target protein HDAC2, blue fluorescence (DAPI staining) for the nucleus. Data are presented as the mean ± standard deviation in the corresponding histogram. ∗ P < 0.05 vs. the control group. Control: rabbit tracheal stenosis model without treatment; NS: rabbit tracheal stenosis model treated with penicillin; ERY: rabbit tracheal stenosis model treated with erythromycin; Budesonide: rabbit tracheal stenosis model treated with budesonide; Vorinostat: rabbit tracheal stenosis model treated with vorinostat. HDAC2, histone deacetylase-2; IF, immunofluorescence.

Article Snippet: Erythromycin enteric-coated tablets (H42021990, Yichang Humanwell Pharmaceutical Co., LTD.); Vorinostat Capsules (180509, Beijing Hengrui Kangda Medical Science and Technology Development Co., Ltd.); Budesonide (AstraZeneca 8339000); Rabbit Anti-Collagen III Polyclonal Antibody (bs-10423R, Bioss); Rabbit Anti-Collagen I Polyclonal Antibody (bs-0549R, Bioss); Rabbit Anti-HDAC2 Polyclonal Antibody (bs-1813R, Bioss); Rabbit VEGF ELISA kit (MM-021001); Rabbit TGF- β 1 ELISA kit (MM-3684001); Rabbit Polyclonal Anti-VEGF (bs-1313R, Bioss, 1/500–1/2000); Rabbit Polyclonal Anti-TGF β 1 (bs-0086R, Bioss, 1/500–1/2000); Rabbit monoclonal Anti-IL-8 (ab34100, abcam, 1/1000); Rabbit Polyclonal Anti-HDAC2 (OmnimAbs, OM105905, 1/500–1/2000); fluorescence microscope (CKX53, OLYMPUS); Microplate Reader (RT-6100, Rayto); Protein vertical electrophoresis instrument (DYY-6C, Beijing 61 instrument factory); Ultra High Sensitivity Chemiluminescence Imaging System (Chemi DocTM XRS+, Bio-Rad Shanhhai Laboratories).

Techniques: Expressing, Immunofluorescence, Fluorescence, Staining, Standard Deviation, Histone Deacetylase Assay

Journal: iScience

Article Title: Alphaherpesvirus manipulates retinoic acid metabolism for optimal replication

doi: 10.1016/j.isci.2024.110144

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-HDAC2 , Proteintech , Cat# 12922-3-AP RRID: AB_2118516.

Techniques: Recombinant, Transfection, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Software

Journal: Cell chemical biology

Article Title: HDAC3 and HDAC8 PROTAC dual degrader reveals roles of histone acetylation in gene regulation

doi: 10.1016/j.chembiol.2023.07.010

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Rabbit polyclonal anti-HDAC2 , Santa Cruz Biotechnology , Cat# sc-7899, RRID: AB_2118563.

Techniques: Recombinant, Software

Journal: iScience

Article Title: Modification of lysine-260 2-hydroxyisobutyrylation destabilizes ALDH1A1 expression to regulate bladder cancer progression

doi: 10.1016/j.isci.2023.108142

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal anti-HDAC2 , Proteintech , Cat #:12922; RRID:AB_2934877.

Techniques: Virus, Recombinant, Activity Assay, SYBR Green Assay, Software

Journal: Research

Article Title: ATP5O Hypo-crotonylation Caused by HDAC2 Hyper-Phosphorylation Is a Primary Detrimental Factor for Downregulated Phospholipid Metabolism under Chronic Stress

doi: 10.34133/2022/9834963

Figure Lengend Snippet: Upregulated HDAC2 phosphorylation is connected with downregulated ATP5O crotonylation in CS mice. (a, b) TMT-labeled phosphoproteomics showed that at the threshold of ≥1.2 or ≤ 0.833 , there are 117 downregulated and 60 upregulated phosphorylated proteins. (c) KEGG pathway showed that multiple pathways are lipid metabolism related. (d) We selected three deacetylases—HDAC2, KAT7, and CREBBP—to verify their relationship with ATP5O. Western blot showed that inhibition of HDAC2 phosphorylation at S424 with THP (CYGRKKRRQRRRSDSEDEGEGGRR) significantly decreased ATP5O, whereas inhibition of KAT7 phosphorylation at S102 with TKP (CYGRKKRRQRRRSSGSETEQVVDF) or CREBBP phosphorylation at S2361 and S2363 with TCP (CYGRKKRRQRRRQSQPPHSSPSPR) did not change ATP5O protein level at all. (e, f) We generated an ATP5O-S424 phosphorylation-specific antibody with a phosphorylated peptide SDS (phospho) EDEGEGGRR and verified the specificity through two ways. (e) Different amounts of non- and S424-phosphorylated HDAC2 peptide were loaded onto a dot blot PVDF membrane and detected by the HDAC2-S424 P antibody. Only phosphorylated HDAC2 peptide was detected in a dose-dependent manner. (f) When HDAC2-S424 P antibody was preblocked by the phosphorylated HDAC2 peptide, significantly less amount of endogenous ATP5O-K51 cr was detected. (g) Western blot showed that THP treatment significantly decreased HDAC2-S424 P (p-HDAC2) level and upregulated ATP5O-K51 cr (ATP5O cr ) level, whereas did not change KAT7-S102 P (p-KAT) level. Besides, THP treatment did not alter the level of HDAC1-S421 p at all, which shared the highest similarity with THP (Supp. Figure ). (h, i) By live imaging in Sf9 cells, when only ATP5O-WT-EGFP titer was added into sf9 culture media, as time progressed, green fluorescence rapidly increased; in contrast, when both ATP5O-EGFP and HDAC2-WT-TagRFP titers were added, as time went by, the red fluorescence (HDAC2) quickly increased, whereas the increment extent of green fluorescence (ATP5O) was significantly slowed down. (j) Coimmunoprecipitation and western blot showed that HDAC2 significantly interacted with ATP5O but rarely interacted with ATP5A1 or ATP5B. (k) GST-pulldown was carried out to verify the direct binding between ATP5O and HDAC2. GST-ATP5O and his-tagged HDAC2-TagRFP were cloned, expressed, and purified in E.coli (From left, the fourth and fifth lane, red asterisk-labeled) and sf9 cells (From left, the third lane, red asterisk-labeled), respectively. SDS-PAGE showed that GST-ATP5O could significantly pull down HDAC2-TagRFP (From left, the sixth lane, red asterisk-labeled). L-N. Western blot showed that when a fixed amount of ATP5O-WT and increasing amounts of HDAC2-WT were cotransfected into 293 T cells, ATP5O crotonylation level rapidly decreased (l, n); in contrast, when a fixed amount of ATP5O-WT and increasing amounts of HDAC2-S424A (a nonphosphorylable mutant) were cotransfected into 293 T cells, the decrement extent of ATP5O crotonylation level became much smaller (m, n). (o–s) When HDAC2-Flag and ATP5O-StrepII were cotransfected into 293 T cells at an increasing dosage, ATP5O crotonylation (o, p) and ATP levels both increased (q); accordingly, FAM126A and STAT5A levels also increased in a dose-dependent manner (r, s). (t–u) Western blot showed that compared with ATP5O-transfected 293 T cells, HDAC2 and ATP5O cotransfected cells had decreased ATP5O crotonylation, whereas inhibition of proteasome activity with MG132 significantly recovered ATP5O crotonylation level. (v, w) Western blot showed that in an in vitro ubiquitination assay with the ATP5O-E3 ubiquitin ligase complex immunoprecipitated from CS ovaries, the ATP5O ubiquitination level was significantly higher than the level in control ovaries. X. Model: ATP5O crotonylation is resistant to ubiquitination, whereas ATP5O-K51 cr decrotonylated by HDAC2 is susceptible to ubiquitination and protease degradation. Tubulin or actin was used as a loading control. * indicates p < 0.05 , *** indicates p < 0.001 , **** indicates p < 0.0001 .

Article Snippet: Other primary antibodies: Mouse monoclonal anti-GAPDH (Cat#: 30201ES60; YEASEN, Shanghai, China); mouse monoclonal anti- β -Actin (Cat#: A5316-100; Sigma, MS, USA); Mouse monoclonal anti- β -Tubulin (Cat#: sc-5274; Santa Cruz, TX, USA); Mouse monoclonal anti-alpha Tubulin (Acetyl Lys40) (cat#: bsm-33235 M; Bioss, Beijing, China); Rabbit anti-Transferrin polyclonal antibody (Cat#: 17435-1-ap; Proteintech, Chicago, USA); Rabbit anti-ATP5O polyclonal antibody (Cat#: D126152; BBI Life science, Shanghai, China); Rabbit anti-ATP5A1 polyclonal antibody (Cat#: D154243; BBI Life science, Shanghai, China); Rabbit anti-ATPB polyclonal antibody (Cat#: A5286; Selleckchem, Shanghai, China); Mouse anti-COX4L1 monoclonal antibody (Cat#: D190618; BBI Life science, Shanghai, China); Anti-AKT (Ab-129) Rabbit polyclonal antibody (Cat#: D151616-0100; BBI Life science, Shanghai, China); Rabbit anti-Phosphpho AKT (Ser473, Cat# 4060, Cell Signaling Technology); Rabbit anti-Phosphpho RPS6 (Ser235/236, Cat#: D155178; BBI Life science, Shanghai, China); Rabbit anti-STAT5A polyclonal antibody (Cat#: D220085; BBI Life science, Shanghai, China); Rabbit anti-FAM126A polyclonal antibody (Cat#: bs-11554R; Bioss, Beijing, China); Rabbit anti-PTDSS1 polyclonal antibody (Cat#: BS-19583R; Bioss, Beijing, China); Rabbit anti-HDAC2 polyclonal antibody (Cat#: 12922-3-ap, Proteintech, Chicago, USA); Mouse monoclonal anti-strep II Tag (Cat#: YFMA0054, Yifeixue, Nanjing, China); Mouse monoclonal anti-flag Tag (Cat#: D190828, BBI Life science, Shanghai, China); Rabbit anti-HDAC1-S421 p polyclonal antibody (Cat#: AP59578, Abcepta, San Diego, CA, USA); Rabbit anti-Ubinuclein polyclonal antibody (Cat#: DF8873, Affinity Biosciences, Melbourne, Australia); Fibrinogen alpha chain (FGA) Rabbit pAb (Cat#: A1453, ABclonal, Wuhan, Hubei, China); Albumin Polyclonal antibody (Cat#: 16475-1-AP; Proteintech, Rosemont, IL, USA); AffiniPure Goat Anti-Mouse IgG (H + L) (Jackson, Cat#: 115-005-003, West Grove, PA, USA); Rabbit anti-ATP5O-K51 cr rabbit polyclonal antibody was made against ASK (crotonyl) EKKLDQVEKELLC by YuBo BioTech and purified by affinity purification; Rabbit anti-HDAC2-S424 p ,and anti-KAT7-S102 p polyclonal antibody was made against SDS(phospho)EDEGEGGRRC and CPLRQTRSSGS (phospho) ET by Zoonbio BioTech and purified by affinity purification.

Techniques: Phospho-proteomics, Labeling, Western Blot, Inhibition, Generated, Dot Blot, Membrane, Imaging, Fluorescence, Binding Assay, Clone Assay, Purification, SDS Page, Mutagenesis, Transfection, Activity Assay, In Vitro, Ubiquitin Proteomics, Immunoprecipitation, Control

Journal: Research

Article Title: ATP5O Hypo-crotonylation Caused by HDAC2 Hyper-Phosphorylation Is a Primary Detrimental Factor for Downregulated Phospholipid Metabolism under Chronic Stress

doi: 10.34133/2022/9834963

Figure Lengend Snippet: Correcting HDAC2 phosphorylation rescued abnormal lipid metabolism. (a, b) Western blot showed that inhibition of HDAC2 activity (phosphorylation) by a peptide THP (CYGRKKRRQRRRSDSEDEGEGGRR) significantly reduced the level of p-HDAC2, whereas ATP5O crotonylation and gross ATP5O levels significantly increased. (c, d) ATP levels in CS ovaries (c) and livers (d) were significantly lower than control, whereas THP treatment significantly recovers the decreased ATP levels close to control. (e) Mating test showed that the first litter size was significantly lower than control, while THP injection recovered the decreased litter size close to control. (f–h) Lipid metabolomes were analyzed for the female mouse serum of control, CS, and CS + THP group; six repeats per group. PCA plot showed that the metabolites in the CS group are completely separate from control group, with the CS + THP group partially overlapping with the control group (f). Heat map showed that at a threshold of CS + THP vs. CS ≥ 1.5 or ≤ 0.67 , there were 91 differentially upregulated or downregulated lipid metabolites (DULMs or DDLMs) that were recovered close to the control (g). And the beneficial lipid form—PE, PA, and PC—covered the second largest percentage (30 of 91, 32.97%) (g, h). (i) Representative differentially downregulated PEs, which were beneficial for physical health, are shown. (j, k) Western blot showed that STAT5A level significantly decreased in CS group, whereas THP treatment significantly recovered the STAT5A level of CS group close to control group. Tubulin was used as a loading control. * indicates p < 0.05 , ** indicates p < 0.01 , *** indicates p < 0.001 , **** indicates p < 0.0001 .

Article Snippet: Other primary antibodies: Mouse monoclonal anti-GAPDH (Cat#: 30201ES60; YEASEN, Shanghai, China); mouse monoclonal anti- β -Actin (Cat#: A5316-100; Sigma, MS, USA); Mouse monoclonal anti- β -Tubulin (Cat#: sc-5274; Santa Cruz, TX, USA); Mouse monoclonal anti-alpha Tubulin (Acetyl Lys40) (cat#: bsm-33235 M; Bioss, Beijing, China); Rabbit anti-Transferrin polyclonal antibody (Cat#: 17435-1-ap; Proteintech, Chicago, USA); Rabbit anti-ATP5O polyclonal antibody (Cat#: D126152; BBI Life science, Shanghai, China); Rabbit anti-ATP5A1 polyclonal antibody (Cat#: D154243; BBI Life science, Shanghai, China); Rabbit anti-ATPB polyclonal antibody (Cat#: A5286; Selleckchem, Shanghai, China); Mouse anti-COX4L1 monoclonal antibody (Cat#: D190618; BBI Life science, Shanghai, China); Anti-AKT (Ab-129) Rabbit polyclonal antibody (Cat#: D151616-0100; BBI Life science, Shanghai, China); Rabbit anti-Phosphpho AKT (Ser473, Cat# 4060, Cell Signaling Technology); Rabbit anti-Phosphpho RPS6 (Ser235/236, Cat#: D155178; BBI Life science, Shanghai, China); Rabbit anti-STAT5A polyclonal antibody (Cat#: D220085; BBI Life science, Shanghai, China); Rabbit anti-FAM126A polyclonal antibody (Cat#: bs-11554R; Bioss, Beijing, China); Rabbit anti-PTDSS1 polyclonal antibody (Cat#: BS-19583R; Bioss, Beijing, China); Rabbit anti-HDAC2 polyclonal antibody (Cat#: 12922-3-ap, Proteintech, Chicago, USA); Mouse monoclonal anti-strep II Tag (Cat#: YFMA0054, Yifeixue, Nanjing, China); Mouse monoclonal anti-flag Tag (Cat#: D190828, BBI Life science, Shanghai, China); Rabbit anti-HDAC1-S421 p polyclonal antibody (Cat#: AP59578, Abcepta, San Diego, CA, USA); Rabbit anti-Ubinuclein polyclonal antibody (Cat#: DF8873, Affinity Biosciences, Melbourne, Australia); Fibrinogen alpha chain (FGA) Rabbit pAb (Cat#: A1453, ABclonal, Wuhan, Hubei, China); Albumin Polyclonal antibody (Cat#: 16475-1-AP; Proteintech, Rosemont, IL, USA); AffiniPure Goat Anti-Mouse IgG (H + L) (Jackson, Cat#: 115-005-003, West Grove, PA, USA); Rabbit anti-ATP5O-K51 cr rabbit polyclonal antibody was made against ASK (crotonyl) EKKLDQVEKELLC by YuBo BioTech and purified by affinity purification; Rabbit anti-HDAC2-S424 p ,and anti-KAT7-S102 p polyclonal antibody was made against SDS(phospho)EDEGEGGRRC and CPLRQTRSSGS (phospho) ET by Zoonbio BioTech and purified by affinity purification.

Techniques: Phospho-proteomics, Western Blot, Inhibition, Activity Assay, Control, Injection

Journal: Research

Article Title: ATP5O Hypo-crotonylation Caused by HDAC2 Hyper-Phosphorylation Is a Primary Detrimental Factor for Downregulated Phospholipid Metabolism under Chronic Stress

doi: 10.34133/2022/9834963

Figure Lengend Snippet: Model: chronic stress caused aberrant level of two types of PTMs, which reduce cytoplasmic ATP level and consequently reduced beneficial phospholipids. Left: Under chronic stress (CS), HDAC2-S424 p was upregulated, which resulted in downregulated ATP5O-K51 cr . Downregulated ATP5O-K51 cr consequently conduced to reduced gross ATP5O level, which finally caused decreased cytoplasmic ATP level and significantly decreased oocyte quality, fertility, and beneficial phospholipids. Middle: TAC is a competitive peptide that was able to specifically downregulate ATP5O-K51 cr . Downregulated ATP5O-K51 cr consequently conduced to reduced gross ATP5O level and cytoplasmic ATP level, which finally significantly decreased oocyte quality, fertility, and beneficial phospholipids. Therefore, TAC treatment could partially recapitulate the decreased fertility and phospholipid in CS mice. Right: THP is a competitive peptide that was able to specifically downregulate the abnormally high level of HDAC2-S424 p in CS mice, which in turn recovered downregulated ATP5O-K51 cr , gross ATP5O level, and cytoplasmic ATP level close to control. Recovered ATP level finally recovered oocyte quality, fertility, and phospholipid metabolism in CS mice, Therefore, THP treatment could partially rescue the decreased fertility and phospholipid in CS mice.

Article Snippet: Other primary antibodies: Mouse monoclonal anti-GAPDH (Cat#: 30201ES60; YEASEN, Shanghai, China); mouse monoclonal anti- β -Actin (Cat#: A5316-100; Sigma, MS, USA); Mouse monoclonal anti- β -Tubulin (Cat#: sc-5274; Santa Cruz, TX, USA); Mouse monoclonal anti-alpha Tubulin (Acetyl Lys40) (cat#: bsm-33235 M; Bioss, Beijing, China); Rabbit anti-Transferrin polyclonal antibody (Cat#: 17435-1-ap; Proteintech, Chicago, USA); Rabbit anti-ATP5O polyclonal antibody (Cat#: D126152; BBI Life science, Shanghai, China); Rabbit anti-ATP5A1 polyclonal antibody (Cat#: D154243; BBI Life science, Shanghai, China); Rabbit anti-ATPB polyclonal antibody (Cat#: A5286; Selleckchem, Shanghai, China); Mouse anti-COX4L1 monoclonal antibody (Cat#: D190618; BBI Life science, Shanghai, China); Anti-AKT (Ab-129) Rabbit polyclonal antibody (Cat#: D151616-0100; BBI Life science, Shanghai, China); Rabbit anti-Phosphpho AKT (Ser473, Cat# 4060, Cell Signaling Technology); Rabbit anti-Phosphpho RPS6 (Ser235/236, Cat#: D155178; BBI Life science, Shanghai, China); Rabbit anti-STAT5A polyclonal antibody (Cat#: D220085; BBI Life science, Shanghai, China); Rabbit anti-FAM126A polyclonal antibody (Cat#: bs-11554R; Bioss, Beijing, China); Rabbit anti-PTDSS1 polyclonal antibody (Cat#: BS-19583R; Bioss, Beijing, China); Rabbit anti-HDAC2 polyclonal antibody (Cat#: 12922-3-ap, Proteintech, Chicago, USA); Mouse monoclonal anti-strep II Tag (Cat#: YFMA0054, Yifeixue, Nanjing, China); Mouse monoclonal anti-flag Tag (Cat#: D190828, BBI Life science, Shanghai, China); Rabbit anti-HDAC1-S421 p polyclonal antibody (Cat#: AP59578, Abcepta, San Diego, CA, USA); Rabbit anti-Ubinuclein polyclonal antibody (Cat#: DF8873, Affinity Biosciences, Melbourne, Australia); Fibrinogen alpha chain (FGA) Rabbit pAb (Cat#: A1453, ABclonal, Wuhan, Hubei, China); Albumin Polyclonal antibody (Cat#: 16475-1-AP; Proteintech, Rosemont, IL, USA); AffiniPure Goat Anti-Mouse IgG (H + L) (Jackson, Cat#: 115-005-003, West Grove, PA, USA); Rabbit anti-ATP5O-K51 cr rabbit polyclonal antibody was made against ASK (crotonyl) EKKLDQVEKELLC by YuBo BioTech and purified by affinity purification; Rabbit anti-HDAC2-S424 p ,and anti-KAT7-S102 p polyclonal antibody was made against SDS(phospho)EDEGEGGRRC and CPLRQTRSSGS (phospho) ET by Zoonbio BioTech and purified by affinity purification.

Techniques: Control