HA723022

HDAC5 Recombinant Rabbit Monoclonal Antibody [PSH08-70]

cat.: HA723022

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	IHC-P, IF-Cell, IP, ChIP
Clonality:	Monoclonal
Clone number:	PSH08-70

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 122 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within human HDAC5 aa 171-220.
Positive control:	Human stomach tissue, mouse stomach tissue, rat stomach tissue, HepG2, NIH/3T3, C6.
Subcellular location:	Nucleus, Cytoplasm.
Recommended Dilutions: IHC-P IF-Cell IP ChIP	1:1,000 1:100 1-2μg/sample Use 0.5~2 μg for 25 μg of chromatin.
Uniprot #:	SwissProt: Q9UQL6 Human \| Q9Z2V6 Mouse Entrez Gene: 84580 Rat
Alternative names:	Antigen NY CO 9 Antigen NY-CO-9 HD5 HDAC 5 HDAC5 HDAC5_HUMAN Histone deacetylase 5 NY CO 9

Images

	Fig1: Immunohistochemical analysis of paraffin-embedded human stomach tissue with Rabbit anti-HDAC5 antibody (HA723022) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA723022) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig2: Immunohistochemical analysis of paraffin-embedded mouse stomach tissue with Rabbit anti-HDAC5 antibody (HA723022) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA723022) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig3: Immunohistochemical analysis of paraffin-embedded rat stomach tissue with Rabbit anti-HDAC5 antibody (HA723022) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA723022) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Fig4: Immunocytochemistry analysis of HepG2 cells labeling HDAC5 with Rabbit anti-HDAC5 antibody (HA723022) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HDAC5 antibody (HA723022) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Fig5: Immunocytochemistry analysis of NIH/3T3 cells labeling HDAC5 with Rabbit anti-HDAC5 antibody (HA723022) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HDAC5 antibody (HA723022) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

	Fig6: Immunocytochemistry analysis of C6 cells labeling HDAC5 with Rabbit anti-HDAC5 antibody (HA723022) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-HDAC5 antibody (HA723022) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
	Fig7: HDAC5 was immunoprecipitated from 0.2 mg NIH/3T3 cell lysate with HA723022 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA723022 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: NIH/3T3 cell lysate (input) Lane 2: HA723022 IP in NIH/3T3 cell lysate Lane 3: Rabbit IgG instead of HA723022 in NIH/3T3 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 5 seconds; ECL: K1801
	Fig8: Chromatin immunoprecipitations were performed with cross-linked chromatin from NIH/3T3 cells with HDAC5 (HA723022) or Normal Rabbit IgG according to the ChIP protocol. The enriched DNA was quantified by real-time PCR using indicated primers. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

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