Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Mammal

Cited:

Mouse

Applications

Validated:

Western Blot, Immunocytochemistry/ Immunofluorescence, Chromatin Immunoprecipitation (Negative), Dot Blot

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all Histone H4 Primary Antibodies »

Product Specifications

Immunogen

Synthetic acetylated peptide surrounding Lysine 16 of human Histone H4 [Swiss Prot P62805].

Reactivity Notes

Predicted to react with most mammal species.

Modification

ac Lys16

Localization

Nucleus. Chromosome.

Marker

H4k16ac

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Histone H4 [ac Lys16] Antibody - BSA Free

Western Blot: Histone H4 [ac Lys16] AntibodyBSA Free [NB21-2077]

Western Blot: Histone H4 [ac Lys16] AntibodyBSA Free [NB21-2077]

Western Blot: Histone H4 [ac Lys16] Antibody [NB21-2077] - Western Blot using NB21-2077 of porcine whole ovarian homogenate. Image submitted via verified customer review.
Immunocytochemistry/ Immunofluorescence: Histone H4 [ac Lys16] Antibody - BSA Free [NB21-2077]

Immunocytochemistry/ Immunofluorescence: Histone H4 [ac Lys16] Antibody - BSA Free [NB21-2077]

Immunocytochemistry/Immunofluorescence: Histone H4 [ac Lys16] Antibody [NB21-2077] - Histone H4 K16ac antibody was tested in HeLa cells with FITC (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and Dylight 550 (red).
Western Blot: Histone H4 [ac Lys16] AntibodyBSA Free [NB21-2077]

Western Blot: Histone H4 [ac Lys16] AntibodyBSA Free [NB21-2077]

Western Blot: Histone H4 [ac Lys16] Antibody [NB21-2077] - WB analysis of H4K16ac in 1) HeLa histone prep lysate and 2) NIH 3T3 histone prep lysate.
Dot Blot: Histone H4 [ac Lys16] Antibody [NB21-2077] - Dot blot analysis of H4K16ac in picomoles of protein.
Histone H4 [ac Lys16] Antibody [NB21-2077]

Applications for Histone H4 [ac Lys16] Antibody - BSA Free

Application
Recommended Usage

Dot Blot

1:1000

Immunocytochemistry/ Immunofluorescence

1:100

Western Blot

1:1000
Application Notes
This H4K16ac antibody is useful for Western Blot, Immunocytochemistry/Immunofluorescence, and Dot Blot. In Western Blot, a band is seen ~13kDa in HeLa histone prep and NIH 3T3 histone prep. In ICC/IF, nuclear staining was observed in HeLa cells. This antibody is not applicable for Chromatin Immunoprecipitation.

Reviewed Applications

Read 1 review rated 5 using NB21-2077 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS and 30% Glycerol

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

0.62 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Histone H4

Chromatin is the arrangement of DNA and proteins in which chromosomes are formed. Correspondingly, chromatin is formed from nucleosomes, which are comprised of a set of four histone proteins (H2A, H2B, H3, H4) wrapped with DNA. Chromatin is a very dynamic structure in which numerous post-translational modifications work together to activate or repress the availability of DNA to be copied, transcribed, or repaired. These marks decide which DNA will be open and commonly active (euchromatin) or tightly wound to prevent access and activation (heterochromatin). Common histone modifications include methylation of lysine and arginine, acetylation of lysine, phosphorylation of threonine and serine, and sumoylation, biotinylation, and ubiquitylation of lysine. In particular, acetylation of H4 Lys16 (H4 K16Ac) plays a role in transcriptional activation and DNA repair. The protein MOF (MYST1) is the primary enzyme to acetylate K16, along with Gcn5, Esal, ATF2, and Sas2. Depletion of MOF reduces K16 acetylation, causing cell cycle arrests, chromosomal aberrations, and a poor DNA damage response.

Alternate Names

H4, H4/M, H4FM, H4M, HIST1H4I, HIST4H4, Histone Cluster 1 H4, Histone Cluster 1 H4i, Histone Cluster 4 H4, H4K16ac

Entrez Gene IDs

121504 (Human); 320332 (Mouse)

Gene Symbol

H4-16

UniProt

P62805 (Human), P62806 (Mouse)

Additional Histone H4 Products

Product Documents for Histone H4 [ac Lys16] Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for Histone H4 [ac Lys16] Antibody - BSA Free

EpiPlus antibodies are produced in collaboration with Rockland Immunochemicals Inc.

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Related Research Areas

Epigenetics
Notch Pathway

Citations for Histone H4 [ac Lys16] Antibody - BSA Free

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  • Histone H4 [ac Lys16] Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Ovary tissue
    Species: Pig
    Verified Customer | Posted 11/17/2016
    H4K16AC on porcine whole ovarian homogenate
    Sample Information: Cell Line or Tissue: Ovary Species: Pig Treatment: None. Run on control tissue Controls: Positive Control: None Negative Control: Primary only, secondary only, and secondary with IgG all negative Loading Control (please attach additional images if applicable): Ponceau S stain Total Protein Loaded: 40 ug/well Electrophoresis: Gel Percentage: Gradient, 4-20% Voltage: 50V then 90V Time: 5 min then 1 hr Membrane Transfer: Method (Submersion/Semi-dry): Semi-dry using iBlot2 system Membrane Type (PVDF/Nitrocellulose): Nitrocellulose Time: 7 min (iBlot2 Protocol 0) Voltage: Click here to enter text. Blocking: Blocking Solution: 5% BSA in.2% PBST Time: 1.5 hours Primary Antibody: Dilution: 1:1000 Diluent Buffer: 5% BSA in.2% PBST Incubation Time: overnight Incubation Temperature: 4°C Washing Conditions: Wash Solution:.2% PBST Time and Repetitions: 3X, 10 minutes each Secondary Antibody Manufacturer and Catalog #: Cell Signaling, #7074 Secondary description: goat anti-rabbit Dilution: 1:1000 Diluent Buffer: 5% BSA in.2% PBST Incubation Time: 1 hour Incubation Temperature: room temperature Detection Method: Detection: SignalFire ECL reagent (Cell Signaling) Procedure: ECL on blot 3 min Development Time: 5 min Molecular weight of band(s): 13 kDa
    Histone H4 [ac Lys16] Antibody - BSA Free NB21-2077

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Protocols

View specific protocols for Histone H4 [ac Lys16] Antibody - BSA Free (NB21-2077):

Histone H4 [ac Lys16] Antibody:
Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 10 ug of histone preps per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.


Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

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Associated Pathways

Notch Signaling Pathways Notch Signaling Pathway Thumbnail