51-5100

antibody from Invitrogen Antibodies

Targeting: HDAC2 KDAC2, RPD3, YAF1

Provider product page for 51-5100

Western blot

ELISA

Immunocytochemistry

Immunoprecipitation

Immunohistochemistry

Flow cytometry

Chromatin Immunoprecipitation

Other assay

Antibody data

Antibody Data
Antigen structure
References [26]
Comments [0]
Validations
Immunocytochemistry [2]
Flow cytometry [1]
Other assay [18]

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Product number: 51-5100 - Provider product page
Provider: Invitrogen Antibodies
Product name: HDAC2 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Reactivity: Human, Mouse
Host: Rabbit
Isotype: IgG
Vial size: 100 µg
Concentration: 0.25 mg/mL
Storage: -20°C

HDAC2 protein structure - 51-5100 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of HDAC2 was done on 70% confluent log phase A431 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with HDAC2 Rabbit Polyclonal Antibody (Product # 51-5100) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing nuclear localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of HDAC2 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with HDAC2 Polyclonal Antibody (Product # 51-5100) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 1 HDAC2 physically interacts with PP2A. a To confirm a physical interaction between PPP2R5A, the regulatory subunit of PP2A, and HDAC2 in NRVCs, primary cultured cardiomyocytes, 2 mg of protein was immunoprecipitated with an anti-PPP2R5A antibody, and the HDAC2 interaction was visualized with an anti-HDAC2 antibody. b-d The catalytic subunit of PP2A, PPP2CA, also bound to HDAC2. To check the interaction in H9c2 cells, a cardiomyoblast cell line, HDAC2-V5 and PPP2CA-myc were transiently transfected, and cells were lysed with 1% NP-40 lysis buffer. Immunoprecipitation was performed with anti-V5 antibody followed by immunoblotting with anti-Myc antibody. c To check the endogenous interaction in NRVCs, primary cultured cardiomyocytes were lysed in 1% NP-40 lysis buffer. Immunoprecipitation was performed with anti-HDAC2 antibody followed by immunoblotting with anti-PPP2CA antibody. d PPP2CA physically interacted with HDAC2 in mouse heart. Two milligrams of heart lysates were immunoprecipitated with anti-PPP2CA. HDAC2 successfully precipitated with PPP2CA

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6. MyoD and HDAC2 repress myogenic late genes at early times of differentiation. ( A ) Time course of gene expression demonstrating that bypassing MyoD by differentiating B22 cells with myogenin/Mef2D1b resulted in premature late gene activation. mRNA levels were normalized to EF1alpha levels. ( B ) ChIP time course indicating that HDAC2 did not bind to late gene regulatory sequences in myogenin/Mef2D1b differentiated B22 cells where MyoD is not expressed. The binding in the myogenin/Mef2D expressing cells at time 0 was normalized to 1. Data are presented as the average of three or more independent experiments +- standard deviation.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 6 ZBTB32 represses Eomes and Cd27 gene expression in CD8 + T cells by recruiting histone deacetylases 1 and 2. WT or Zbtb32 -/- P14 splenocytes were transferred into recipients, which were then infected with LCMV-Armstrong. At days 6, 8 and 10 post-infection, P14 cells were isolated and pooled from three mice per genotype for RNA isolation; chromatin was prepared at day 7 post-infection. (a) Eomes and Cd27 mRNA levels were examined by quantitative RT-PCR relative to Actb mRNA. (b) Schematic of Eomes and Cd27 gene loci showing position of specific (Amplicon 1; Amp1) and non-specific (Amplicon 2; Amp2) primers. In each case, Amp1 corresponds to putative ZBTB32 binding site. (c) The enrichment of ZBTB32 on Eomes and Cd27 genes by chromatin immunoprecipitation (ChIP). (d) The enrichment of HDAC1 and HDAC2 on Eomes and Cd27 genes by ChIP. (e) ChIP for Pol II, p300 or modified histone H3 at the Eomes and Cd27 loci. Data are a compilation of three independent experiments; error bars represent the SEM. Iso; isotype control antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. 4 Interaction candidates are coexpressed with TBX2 in the pulmonary mesenchyme and interact in HEK293 cells. a Co-immunofluorescence analysis of candidate interaction partners (red) and TBX2 (green) on frontal sections of the right lung of E14.5 Tbx2 cre/ + embryos. Antigens are color-coded and nuclei were counterstained with DAPI (blue). Insets or selected regions in overview images are magnified in rows 2,4 and 6. b In situ proximity ligation assay of TBX2 and candidate interaction partners on 10 um frontal sections of E14.5 wildtype and Tbx2 cre/fl mutant lungs. Direct interaction is visualized by small red fluorescent dots. Larger more diffuse orange stains are due to auto-fluorescence of blood cells. Nuclei are counterstained with DAPI (blue). c Western blot analysis of co-immunoprecipitation experiments for verification of TBX2 interaction with candidate proteins on 10% SDS polyacrylamide gels. Detection was performed with an anti-TBX2 primary antibody and developed with chemoluminescence-IHC. Arrows indicate TBX2 bands. Lanes were loaded as follows: No antibody: IP without specific antibody resembling negative IP-control; 5% input: 5% of crude cell extract before precipitation; empty: no protein loaded; IP: co-immunoprecipitate with antibody for specific candidate. Expected molecular weight for TBX2.HA approx. 76.2 kDa

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: VPA but not TSA leads to reduction of HDAC2 protein levels. ( A ) K562 human erythroleukemia cells were treated for 24 h as indicated with the HDAC inhibitors VPA (1.5 mM), TSA (100 nM), butyrate (1.5 mM) or MS-27-275 (5 muM). Amounts of HDAC2 protein were determined by western blot analysis of whole-cell extracts. Actin protein levels were determined to verify equal loading of samples. ( B ) F9 mouse teratocarcinoma or HEK293T human embryonic kidney carcinoma cells were exposed to 1 mM VPA for the indicated periods of time. Protein levels of HDAC2 as well as HDAC1, HDAC3 and actin were determined by western blot analysis. ( C ) The dose-dependent reduction of HDAC2 protein levels was determined in F9 or HEK293T cells after exposure to VPA for 30 or 24 h, respectively. ( D ) Time course analyses in F9 and HEK293T cells confirmed that TSA (100 nM) does not affect the amount of HDAC2 protein. ( E ) Reduction of HDAC2 protein levels after treatment of mice with VPA was tested by western blot analysis of tissue extracts and immunohistochemistry. Similar results were obtained in at least two sets of independent experiments.