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Histone H3 (acetyl K14) Recombinant Antibody

The Hela (treated with Trichostatin A (TSA)(1\u03bcM, 18h)) (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-Histone H3 (acetyl K14) antibody (bsm-54178R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
The Hela (treated with Trichostatin A (TSA)(1\u03bcM, 18h)) (H) cells were fixed with 4% PFA (10 min at r.t.) and then permeabilized with 90% ice-cold methanol for 20 min at -20\u2103,the cells then were incubated in 5%BSA to block non-specific protein-protein interactions (30 min at r.t.), followed by secondary antibody incubation for 40 min at room temperature. Primary Antibody (green):Rabbit Anti-Histone H3 (acetyl K14) antibody (bsm-54178R,1:100); Isotype Control (orange): Rabbit IgG (bs-0295P). Blank control (black): PBS. Acquisition of 20,000 events was performed.
4% Paraformaldehyde-fixed Hela (treated with Trichostatin A (TSA)(1\u03bcM, 18h)) (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Histone H3 (acetyl K14)) monoclonal Antibody, unconjugated (bsm-54178R) 1:100, 90 min at 37\u00b0C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
4% Paraformaldehyde-fixed Hela (treated with Trichostatin A (TSA)(1\u03bcM, 18h)) (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Histone H3 (acetyl K14)) monoclonal Antibody, unconjugated (bsm-54178R) 1:100, 90 min at 37\u00b0C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (acetyl K14) Monoclonal Antibody, Unconjugated (bsm-54178R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (acetyl K14) Monoclonal Antibody, Unconjugated (bsm-54178R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (acetyl K14) Monoclonal Antibody, Unconjugated (bsm-54178R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (acetyl K14) Monoclonal Antibody, Unconjugated (bsm-54178R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (acetyl K14) Monoclonal Antibody, Unconjugated (bsm-54178R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (acetyl K14) Monoclonal Antibody, Unconjugated (bsm-54178R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (acetyl K14) Monoclonal Antibody, Unconjugated (bsm-54178R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (acetyl K14) Monoclonal Antibody, Unconjugated (bsm-54178R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (acetyl K14) Monoclonal Antibody, Unconjugated (bsm-54178R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (acetyl K14) Monoclonal Antibody, Unconjugated (bsm-54178R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (acetyl K14) Monoclonal Antibody, Unconjugated (bsm-54178R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Histone H3 (acetyl K14) Monoclonal Antibody, Unconjugated (bsm-54178R) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-0295G-HRP and DAB (C-0010) staining.
C2C12 (M) treated with Trichostatin A (TSA) (1\u03bcM) for 18 h, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with Histone H3 (acetyl K14) monoclonal antibody, unconjugated (bsm-54178R) at 1:800 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
C2C12 (M) treated with Trichostatin A (TSA) (1\u03bcM) for 18 h, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with Histone H3 (acetyl K14) monoclonal antibody, unconjugated (bsm-54178R) at 1:800 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Chromatin immunoprecipitation was performed using cross-linked chromatin from 5\u00d710\u2076 HeLa cells (treated with 5 mM sodium butyrate for 24 hours) per IP, with 5 \u03bcg of bsm-54178R antibody or 5 \u03bcg of normal rabbit IgG(negative control bs-0295P) , and 36 \u03bcL of Protein A\/G MagBeads per IP. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human GAPDH-intron 2,GAPDH-intron 2 and RPL30 exon 3 regions. The data are presented as enrichment of each sample relative to the total amount of input at each amplicon.
Chromatin immunoprecipitation was performed using cross-linked chromatin from 5\u00d710\u2076 HeLa cells (treated with 5 mM sodium butyrate for 24 hours) per IP, with 5 \u03bcg of bsm-54178R antibody or 5 \u03bcg of normal rabbit IgG(negative control bs-0295P) , and 36 \u03bcL of Protein A\/G MagBeads per IP. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human GAPDH-intron 2,GAPDH-intron 2 and RPL30 exon 3 regions. The data are presented as enrichment of each sample relative to the total amount of input at each amplicon.
Bioss unconjugated antibody vial

Applications

  • WB
  • IHC-P
  • IHC-F
  • IF
  • ICC/IF

Reactivity

  • Human
  • Mouse
  • Rat
Overview
Catalog # bsm-54178r-100uL
Product Name Histone H3 (acetyl K14) Recombinant Antibody
Applications
WB IHC-P IHC-F IF ICC/IF
Reactivity Human, Mouse, Rat
Specifications
Conjugation Unconjugated
Host Rabbit
Source Synthetic peptide of human Histone H3 (acetyl K14) coupled with KLH
Modification Site acetyl K14
Clonality Recombinant
Isotype IgG
Concentration Lot Dependent
Purification Purified by Protein A.
Storage Buffer 0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol
Storage Condition Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID 8350
Swiss Prot P68431
Subcellular location Nucleus
Synonyms Histone H3.1; Histone H3/a; Histone H3/b; Histone H3/c; Histone H3/d; Histone H3/f; Histone H3/h; Histone H3/i; Histone H3/j; Histone H3/k; Histone H3/l; HIST1H3A; H3FA; HIST1H3B; H3FL; HIST1H3C; H3FC; HIST1H3D; H3FB; HIST1H3E; H3FD; HIST1H3F; H3FI; HIST1H3G; H3FH; HIST1H3H; H3FK; HIST1H3I; H3FF; HIST1H3J; H3FJ.
Background Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the 'beads on a string' structure. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine.
Application Dilution
WB 1:300-5000
IHC-P 1:200-400
IHC-F 1:100-500
IF
ICC/IF 1:50-100

Frequently asked questions

Which applications is Histone H3 (acetyl K14) Recombinant Antibody (bsm-54178R) validated for, and at what dilution?

bsm-54178R is validated for WB (1:300-5000); IHC-P (1:200-400); IHC-F (1:100-500); IF; ICC/IF (1:50-100). These are recommended starting dilutions; optimal dilutions should be determined by the end user.

Which species does Histone H3 (acetyl K14) Recombinant Antibody (bsm-54178R) react with?

Tested reactivity: Human, Mouse, Rat.

How should bsm-54178R be stored?

Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles. Storage buffer: 0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol

What immunogen was used to raise Histone H3 (acetyl K14) Recombinant Antibody (bsm-54178R)?

Synthetic peptide of human Histone H3 (acetyl K14) coupled with KLH

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