Histone H4 [Monomethyl Lys20] Antibody

Product Discontinued

Histone H4 [Monomethyl Lys20] Antibody Summary

• Immunogen: Synthetic monomethylated peptide surrounding Lysine 20 of human Histone H4 [Swiss Prot P62805].
• Modification: Monomethyl Lys20
• Localization: Nucleus. Chromosome.
• Marker: H4K20me1
• Predicted Species: Rat (100%), Chicken (100%), Xenopus (100%), Drosophila (100%), Plant (100%). Backed by our 100% Guarantee.
• Clonality: Polyclonal
• Host: Rabbit
• Gene: HIST4H4
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Chromatin Immunoprecipitation 2-5 ug per million cells
  • Dot Blot 1:1000
  • Immunocytochemistry/ Immunofluorescence 1:50
  • Western Blot 1-2 ug/ml
• Application Notes: This Histone H4 K20me1 antibody is useful for Immunocytochemistry/Immunofluorescence ,ChIP, Dot blot, and Western blot where a band is seen ~13kDa.
• Control:

  GAPDH primer
  GAPDH primer
  MYOD1 primer
  MYOD1 primer
  AFM primer
  AFM primer
  alpha Satellite repeat primer
  alpha Satellite repeat primer
  Satellite2 repeat primer
  Satellite2 repeat primer

Reactivity Notes

Human, mouse, and C. elegans. Predicted to react with many species including rat, chicken, Xenopus, Drosophila, and plant based on 100% sequence homology.

Packaging, Storage & Formulations

• Storage: Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
• Buffer: PBS and 30% Glycerol
• Preservative: 0.05% Sodium Azide
• Concentration: 1.12 mg/ml
• Purity: Immunogen affinity purified

Notes

Epi-Plus antibody production in collaboration with Rockland Immunochemicals Inc.

Alternate Names for Histone H4 [Monomethyl Lys20] Antibody

• H4
• H4/A
• H4/B
• H4/C
• H4/D
• H4/E
• H4/G
• H4/H
• H4/I
• H4/J
• H4/K
• H4/M
• H4/N
• H4/O
• H4/p
• H4F2
• H4FA
• H4FB
• H4FC
• H4FD
• H4FE
• H4FG
• H4FH
• H4FI
• H4FJ
• H4FK
• H4FM
• H4FN
• H4FO
• H4K20Me1
• H4M
• HIST1H4A
• HIST1H4B
• HIST1H4C
• HIST1H4D
• HIST1H4E
• HIST1H4F
• HIST1H4H
• HIST1H4I
• HIST1H4J
• HIST1H4K
• HIST1H4L
• HIST2H4
• HIST2H4A
• HIST2H4B
• HIST4H4
• histone 4, H4
• Histone Cluster 1 H4
• Histone Cluster 1 H4i
• Histone Cluster 4 H4
• histone cluster 4, H4
• Histone H4
• MGC24116

Background

SET8 specifically catalyzes the mono-methylation of H4 at K20. Loss of this crucial methylation causes multiple DNA breaks, which instigates a p53-dependent DNA damage response to avoid mitosis and aberrant chromosomal activity. Therefore, this PTM is essential to genome replication and stability through S-phase. In mammalian stem cells, Xist expression blocks the formation of H4K20me1, which is one of the first examples of a direct connection between chromatin and stem cell differentiation.

Limitations

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.

Publications for Histone H4 Antibody (NB21-2088)(2)

Publications using NB21-2088

• Bidne KL. Investigating the ovarian response to endotoxemia. Thesis 1905-07-09 (WB, Porcine)
• Martinez Redondo P. The epigenetic regulation of cell cycle and chromatin dynamic by sirtuins Thesis et al. 2014-01-01 (ICC/IF, IHC-Fr, Mouse)
  
  Details:
  Histone H4 [Monomethyl Lys20] H4K20me1 antibody used for ICC-IF and IHC-Fr applications on samples from WT and SIRT2 KO mice.

Reviews for Histone H4 Antibody (NB21-2088) (2)

Average Rating: 4

(Based on 2 reviews)

We have 2 reviews tested in 2 species: Mouse, Pig.

reviewed by: Verified Customer WB Pig 11/17/2016

Summary

• Application: Western Blot
• Sample Tested: Ovary tissue
• Species: Pig

Comments

• Comments: Sample Information:
       Treatment: None. Run on control tissue
  
  Controls:
       Positive Control: None
       Negative Control: Primary only, secondary only, and secondary with IgG all negative
       Loading Control: Ponceau S stain
  
  Total Protein Loaded: 40 ug/well
  
  Electrophoresis:
  Gel Percentage: Gradient, 4-20%
  Electrophoresis Conditions:
       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)
  
  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

reviewed by: Verified Customer WB Mouse 06/09/2016

Summary

• Application: Western Blot
• Sample Tested: Primary mouse neural progenitors growing in culture
• Species: Mouse

FAQs for Histone H4 Antibody (NB21-2088). (Showing 1 - 1 of 1 FAQs).

1. We have a question about the ChIP result graphs obtained by Epi-Plus antibodies. I am sorry that I could not interpret the results. Could you tell us the interpretation of the results of NB21-2088 and NB21-2089, for example?
   • The graphs for NB21-2088 and NB21-2089 show the relative amount of DNA for each listed target that was co-precipitated with chromatin. Briefly, cell lysates were incubated with magnetic beads and antibodies for specific histone modifications. The beads and antibody will isolate only those histone complexes that have the specific modification (for example, Dimethyl Lys20 for NB21-2089). These histone complexes are also bound to DNA, therefore any DNA sequences that are associated with these complexes will also be isolated. Once the histones/DNA are immuoprecipitated, the DNA is extracted and qPCR is performed to measure the relative abundance of certain genes. For NB21-2089, the graph shows that genes whose protein is highly expressed in HeLa cells are co-precipitated with NB21-2089 much LESS than genes that are silent in these cell types. This is expected because the modification that NB21-2089 targets is associated with gene silencing, therefore you would expect less GAPDH signal, and more MYOD1 signal (which is not expressed in HeLa cells).

Bioinformatics

• Gene Symbol: HIST4H4
• Entrez:
  • 121504(Human)
  • 320332(Mouse)
• Uniprot:
  • P62805(Human)
  • P62806(Mouse)