Histone H4 [Monomethyl Lys20] Antibody (5E10-D8) - BSA Free

Novus Biologicals | Catalog # NBP1-30091

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Key Product Details

Species Reactivity

Human, Mouse, Rat, C. elegans, Drosophila, Mammal, Yeast

Applications

Western Blot, ELISA, Immunocytochemistry/ Immunofluorescence, Chromatin Immunoprecipitation (ChIP), Dot Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 kappa Clone # 5E10-D8

Format

BSA Free
View all Histone H4 Primary Antibodies »

Product Specifications

Immunogen

Branched peptide found between amino acids 10-100 of Histone H4. [UniProt# P62805]

Reactivity Notes

Predicted to react all mammals

Modification

Monomethyl Lys20

Localization

Nucleus

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1 kappa

Theoretical MW

11 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Scientific Data Images for Histone H4 [Monomethyl Lys20] Antibody (5E10-D8) - BSA Free

Chromatin Immunoprecipitation: Histone H4 [Monomethyl Lys20] Antibody (5E10-D8) [NBP1-30091] - HeLa extracts.
Immunocytochemistry/Immunofluorescence: Histone H4 [Monomethyl Lys20] Antibody (5E10-D8) [NBP1-30091] - A431 cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.5% Triton X-100 in PBS for 5 minutes. The cells were incubated with Histone H4 [Monomethyl Lys20] Antibody [5E10-D8] (NBP1-30091) at 1 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.
Western Blot: Histone H4 [Monomethyl Lys20] Antibody (5E10-D8) [NBP1-30091] - Analysis of Histone H4 meIK20 in HeLa histone lysates.
Immunocytochemistry/Immunofluorescence: Histone H4 [Monomethyl Lys20] Antibody (5E10-D8) [NBP1-30091] - Histone H4 [methyl Lys20] staining in Hela cells detected with a DyLight 488 labeled secondary antibody (Green) with Hoechst 33258 nuclear counterstain (Blue).
Dot Blot: Histone H4 [Monomethyl Lys20] Antibody (5E10-D8) [NBP1-30091] - Analysis of Histone H4 me1K20 with 50 pmol or 10 pmol of peptides corresponding to amino acids 12-31 of Histone H4 bearing unmodified, monomethylated, dimethylated or trimethylated K20.

Applications for Histone H4 [Monomethyl Lys20] Antibody (5E10-D8) - BSA Free

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

1:10-1:500

Dot Blot

1:2000

ELISA

1:100-1:2000

Immunocytochemistry/ Immunofluorescence

1:200

Western Blot

1:1000
Application Notes
This Histone H4 [Monomethyl Lys20] (5E10-D8) antibody is useful in Chromatin Immunoprecipitation, Dot Blot, ELISA, Immunocytochemistry/Immunofluorescence and Western blot. In WB a band is seen at ~11 kDa.

Reviewed Applications

Read 1 review rated 5 using NBP1-30091 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein G purified

Formulation

Tris-Glycine and 0.15M NaCl

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1 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

H4Me1K20: Methylation of position-specific lysine residues in histone N termini is a central modification for regulating epigenetic transitions in chromatin. Each methylatable lysine residue can exist in a mono, di, or tri methylated state. Arginine resdiues can also by mono or di methylated.

Alternate Names

H4, H4/M, H4FM, H4M, HIST1H4I, HIST4H4, Histone Cluster 1 H4, Histone Cluster 1 H4i, Histone Cluster 4 H4, H4K20Me1

Entrez Gene IDs

121504 (Human); 320332 (Mouse)

Gene Symbol

H4-16

UniProt

P62805 (Human), P62806 (Mouse), P62804 (Rat)

Additional Histone H4 Products

Product Documents for Histone H4 [Monomethyl Lys20] Antibody (5E10-D8) - BSA Free

Certificate of Analysis

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Product Specific Notices for Histone H4 [Monomethyl Lys20] Antibody (5E10-D8) - BSA Free

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

Customer Reviews for Histone H4 [Monomethyl Lys20] Antibody (5E10-D8) - BSA Free (1)

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  • Name: Anonymous
    Application: Western Blot
    Sample Tested: Horse cell lysate
    Species: Other
    Verified Customer | Posted 09/12/2016
    Analysis of Histone H4 (Lys20) in equine fibroblasts
    Histone H4 [Monomethyl Lys20] Antibody (5E10-D8) - BSA Free NBP1-30091

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Protocols

View specific protocols for Histone H4 [Monomethyl Lys20] Antibody (5E10-D8) - BSA Free (NBP1-30091):

Histone H4 [Monomethyl Lys20] Antibody (5E10-D8):
Procedure Guide for NBP1-30091- Histone H4 [K20-me1] Antibody
Western Blot Protocol
1. Perform SDS-PAGE (4-12% MOPS) on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer
apparatus.
3. Rinse membrane with dH2O and then stain the blot using Ponceau S for 1-2 minutes to access the transfer of
proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations
and locations of molecular weight markers using a pencil.
4. Rinse the blot in TBS for approximately 5 minutes.
5. Block the membrane using 5% BSA in TBS + Tween, 1 hour at RT.
6. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10
minutes each.
7. Dilute the mouse anti-Histone H4 [K20-me1]primary antibody (NBP1-30091) in blocking buffer and incubate 1 hour at room
temperature.
8. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1% Tween] 3 times for 10
minutes each.
9. Apply the diluted mouse-IgG 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 [TBS + 0.1% Tween] 3 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 (Pierce ECL).
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%, provided
it does not interfere with antibody-antigen binding.

(c) 2009 Novus Biologicals - Histone H4 [K20-me1] Antibody - Page 1

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Associated Pathways

Notch Signaling Pathways Notch Signaling Pathway Thumbnail