Hdac9

Manufactured by Santa Cruz Biotechnology

Sourced in United States

HDAC9 is a lab equipment product offered by Santa Cruz Biotechnology. It is a histone deacetylase enzyme that plays a role in the regulation of gene expression.

Automatically generated - may contain errors

Lab products found in correlation

Close spelling variants of this product

Anti-HDAC9

5 protocols using hdac9

Immunoprecipitation of HDAC4, YY1, and HDAC9 from MDA-MB-231 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

MDA-MB-231 cell lysates were prepared by incubating the cells in NP-40 lysis buffer containing protease inhibitor cocktails (1:10,000). Lysates were centrifuged at 12,000 rpm for 10 min at 4°C and incubated with control or specific antibodies for 0.5 h. Add 30 μL protein A/G agarose (Pierce, USA) of each tube at 4°C with constant rotation for 8–12 h. After incubation was performed, the beads were washed 5–6 times by using cold buffer. The precipitated proteins were eluted from the beads by re-suspending the beads in 2× SDS-PAGE loading buffer and boiling for 5 min at 99°C. The boiled immune complexes were subjected to western blotting. The following antibodies were used: HDAC4 (1:100, Proteintech, China), YY1 (1:100, Cell Signaling Technology, USA), HDAC9 (1:100, Santa Cruz Biotechnology, USA), and immunoglobulin G (IgG; 1:100, Cell Signaling Technology, USA).

Guo Q., Wang T., Yang Y., Gao L., Zhao Q., Zhang W., Xi T, & Zheng L. (2020). Transcriptional Factor Yin Yang 1 Promotes the Stemness of Breast Cancer Cells by Suppressing miR-873-5p Transcriptional Activity. Molecular Therapy. Nucleic Acids, 21, 527-541.

+ Open protocol

+ Expand

Modulating HDAC9 and Autophagy in BMMSCs

Check if the same lab product or an alternative is used in the 5 most similar protocols

Small interfering RNA (siRNA) targeting mice BECN1 (Ribobio, China) or HDAC9 (Santa Cruz, USA) were transfected into cells at a final concentration of 50 nM using riboFECT™ CP (Ribobio, China). siNC (Ribobio, China) was used as negative controls. All steps were performed according to the instruction in the riboFECT™ CP kit. The following experiments were performed according to the experiment designed. In detail, siRNA silencing HDAC9 was transfected into BMMSCs to investigate the effects of HDAC9 on osteogenic and adipogenic differentiation of BMMSCs and binding to the promoter of autophagy-related genes. In addition, aged BMMSCs were co-transfected with siHDAC9 and siBECN1 to investigate the hypothetical HDAC9-autophagy axis which may regulate BMMSC lineage differentiation.

Zhang L., Qi M., Chen J., Zhao J., Li L., Hu J., Jin Y, & Liu W. (2020). Impaired autophagy triggered by HDAC9 in mesenchymal stem cells accelerates bone mass loss. Stem Cell Research & Therapy, 11, 269.

+ Open protocol

+ Expand

Western Blot Analysis of Renal Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein expression in cells and renal tissues was detected by Western blotting as described previously44 (link). An equal amount of protein (50 μg) from each sample was separated by SDS-PAGE and transferred to PVDF membranes. After blocking in blocking buffer containing 5% skim milk for 30 min, the membranes were incubated with primary antibodies for HDAC9 (Santa Cruz Biotechnology, Santa Cruz, CA, USA), p-STAT3 (ab76315, Abcam, Cambridge, MA, USA), STAT3 (Abcam), Bcl-2 (Santa Cruz Biotechnology), Bax (Santa Cruz Biotechnology), cleaved Caspase 3 (Abcam), Nephrin (Abcam), Podocin (Abcam) and GAPDH (Cell signaling Technology, Danvers, MA, USA) separately at 4 °C overnight. The membranes were incubated with appropriate HRP-conjugated secondary antibody at room temperature for 1 h and signals were detected by using enhanced chemiluminescence (BioRad, Richmond, CA, USA). The blotting bands were quantified by densitometric analysis using Image J software (National Institutes of Health, Bethesda, MD, USA).

Liu F., Zong M., Wen X., Li X., Wang J., Wang Y., Jiang W., Li X., Guo Z, & Qi H. (2016). Silencing of Histone Deacetylase 9 Expression in Podocytes Attenuates Kidney Injury in Diabetic Nephropathy. Scientific Reports, 6, 33676.

+ Open protocol

+ Expand

Western Blot Analysis of HDAC Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed in TESH buffer [10 mmol/L Tris, 1 mmol/L EDTA, 12 mmol/L thioglycerol (pH 7.7)][19 (link)] with 0.4 M NaCl using three rounds of freeze/thaw; 30 μg of protein were resolved on a 6.5% SDS gel. Proteins were detected using ECL reagents and antibodies raised against HDAC1 (sc-7872, rabbit polyclonal, used at 1:200), HDAC5 (sc-133225, monoclonal, used at 1:100), HDAC6 (sc-11420, rabbit polyclonal, used at 1:200), HDAC7 (sc-74563, monoclonal, used at 1:250), HDAC8 (sc11405, rabbit polyclonal, used at 1:200), HDAC9 (sc 28732, rabbit polyclonal, used at 1:100), and HDAC10 (sc134995, rabbit polyclonal, used at 1:200) all from Santa Cruz (Santa Cruz, CA), AR [monoclonal anti AR antibody (AR441), used at 1:1000] [43 (link)] (a gift of Nancy Weigel, Baylor College of Medicine) and β-actin [rabbit polyclonal antibody from Sigma-Aldrich (A-2066) (used at 1:250)]. Phospho-HER2/ErbB2 (Tyr1248) Antibody #2247 (used at 1:1000) and HER2/ErbB2 Ab #2165 (used at 1:1000) were from Life Technologies. Phospho-EGF Receptor (Tyr1068) Ab # 3777 (used at 1:1000) and EGF Receptor Antibody #2232 (used at 1:1000) were from Life Technologies.

Sun H., Mediwala S.N., Szafran A.T., Mancini M.A, & Marcelli M. (2016). CUDC-101, a Novel Inhibitor of Full-Length Androgen Receptor (flAR) and Androgen Receptor Variant 7 (AR-V7) Activity: Mechanism of Action and In Vivo Efficacy. Hormones & Cancer, 7(3), 196-210.

+ Open protocol

+ Expand

Protein Expression Profiling in Kidney Injury

Check if the same lab product or an alternative is used in the 5 most similar protocols

Antibodies to HDAC4, HDAC5, HDAC7, and HDAC9 were purchased from Santa Cruz Biotechnology (Dallas, TX, United States). Antibodies to NGAL, BMP7, and LC3 were purchased from Abcam Inc (Cambridge, MA United States). Antibodies to p-p53, p53, caspase-3, bcl-2, Bax, PCNA, P-ERK1/2, ERK1/2, E-Cadherin, and β-Actin were purchased from Cell Signaling Technology (Danvers, MA, United States). Antibodies to Atg7 and Beclin-1 were purchased from Arigo Institute (Taiwan, China). Antibodies to GAPDH and α-tubulin were purchased from Proteintech Group Institute (Wuhan, China). Secondary antibodies were purchased from LI-COR Biosciences (Lincoln, NE, United States). Serum creatinine (Scr) reagent kits and Blood urea nitrogen (BUN) reagent kits were purchased from Nanjing Jiancheng Bioengineering Institute (Nanjing, China). All other chemicals were purchased from Sigma (St. Louis, Missouri, United States).

Li J., Yu C., Shen F., Cui B., Liu N, & Zhuang S. (2022). Class IIa histone deacetylase inhibition ameliorates acute kidney injury by suppressing renal tubular cell apoptosis and enhancing autophagy and proliferation. Frontiers in Pharmacology, 13, 946192.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!