Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895)

Overview

  • Product nameAnti-Histone H3 (mono methyl K4) antibody - ChIP Grade
    See all Histone H3 primary antibodies
  • Description
    Rabbit polyclonal to Histone H3 (mono methyl K4) - ChIP Grade
  • SpecificitySpecific for mono-methylated Lysine 4 of histone H3. Does not recognise di- or tri-methyl Lysine 4 nor methylation at Lysine 9.
  • Tested applicationsIP, Flow Cyt, Electron Microscopy, ICC/IF, ICC, ChIP, WB, CHIPseq, IHC-P, ChIP/Chipmore details
  • Species reactivity
    Reacts with: Mouse, Human, Pig, Saccharomyces cerevisiae, Tetrahymena sp., Xenopus laevis, Fruit fly (Drosophila melanogaster), Plasmodium falciparum, Xenopus tropicalis , Candida albicans
    Predicted to work with: Cow, Indian Muntjac, Plants, all Mammals
  • Immunogen

    Synthetic peptide within Human Histone H3 aa 1-100 (mono methyl K4) conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
    (Peptide available as ab1340)

  • Positive control
    • WB: Calf thymus histone preparation and HeLa whole cell extract. ICC/IF: HeLa cells.

Properties

  • FormLiquid
  • Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage bufferpH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • PurityImmunogen affinity purified
  • ClonalityPolyclonal
  • IsotypeIgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab8895 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IP Use at an assay dependent concentration.
Flow Cyt 1/100.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

Electron Microscopy Use at an assay dependent concentration. PubMed: 20543957
ICC/IF Use a concentration of 1 µg/ml.

Works better if cells are fixed with methanol.

ICC Use at an assay dependent concentration.
ChIP Use 2 µg for 25 µg of chromatin.
WB 1/500. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).
CHIPseq Use at an assay dependent concentration. PubMed: 22196736
IHC-P Use a concentration of 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ChIP/Chip Use at an assay dependent concentration.

Target

  • FunctionCore component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
  • Sequence similaritiesBelongs to the histone H3 family.
  • Developmental stageExpressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
  • Post-translational
    modifications
    Acetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8me2s). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me).
    Citrullination at Arg-9 (H3R8ci) and/or Arg-18 (H3R17ci) by PADI4 impairs methylation and represses transcription.
    Asymmetric dimethylation at Arg-18 (H3R17me2a) by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 (H3R8me2s) by PRMT5 is linked to gene repression. Asymmetric dimethylation at Arg-3 (H3R2me2a) by PRMT6 is linked to gene repression and is mutually exclusive with H3 Lys-5 methylation (H3K4me2 and H3K4me3). H3R2me2a is present at the 3' of genes regardless of their transcription state and is enriched on inactive promoters, while it is absent on active promoters.
    Methylation at Lys-5 (H3K4me), Lys-37 (H3K36me) and Lys-80 (H3K79me) are linked to gene activation. Methylation at Lys-5 (H3K4me) facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 (H3K79me) is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are linked to gene repression. Methylation at Lys-10 (H3K9me) is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 (H3S10ph) and acetylation of H3 and H4. Methylation at Lys-5 (H3K4me) and Lys-80 (H3K79me) require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are enriched in inactive X chromosome chromatin.
    Phosphorylated at Thr-4 (H3T3ph) by GSG2/haspin during prophase and dephosphorylated during anaphase. Phosphorylation at Ser-11 (H3S10ph) by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at Ser-11 (H3S10ph) by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-fos and c-jun. Phosphorylation at Ser-11 (H3S10ph), which is linked to gene activation, prevents methylation at Lys-10 (H3K9me) but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 (H3S10ph) by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at Ser-11 (H3S10ph) is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 (H3S28ph) by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation at Thr-7 (H3T6ph) by PRKCBB is a specific tag for epigenetic transcriptional activation that prevents demethylation of Lys-5 (H3K4me) by LSD1/KDM1A. At centromeres, specifically phosphorylated at Thr-12 (H3T11ph) from prophase to early anaphase, by DAPK3 and PKN1. Phosphorylation at Thr-12 (H3T11ph) by PKN1 is a specific tag for epigenetic transcriptional activation that promotes demethylation of Lys-10 (H3K9me) by KDM4C/JMJD2C. Phosphorylation at Tyr-42 (H3Y41ph) by JAK2 promotes exclusion of CBX5 (HP1 alpha) from chromatin.
    Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination (By similarity). Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins.
  • Cellular localizationNucleus. Chromosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • H3 histone family, member J antibody
    • FLJ92264 antibody
    • H3 histone family, member A antibody
    • H3 histone family, member B antibody
    • H3 histone family, member C antibody
    • H3 histone family, member D antibody
    • H3 histone family, member F antibody
    • H3 histone family, member H antibody
    • H3 histone family, member I antibody
    • H3 histone family, member K antibody
    • H3 histone family, member L antibody
    • H3 histone, family 3A antibody
    • H3.3A antibody
    • H3/a antibody
    • H3/b antibody
    • H3/c antibody
    • H3/d antibody
    • h3/f antibody
    • H3/h antibody
    • H3/i antibody
    • H3/j antibody
    • H3/k antibody
    • H3/l antibody
    • H31_HUMAN antibody
    • H3F1K antibody
    • H3F3 antibody
    • H3FA antibody
    • H3FB antibody
    • H3FC antibody
    • H3FD antibody
    • H3FF antibody
    • H3FH antibody
    • H3FI antibody
    • H3FJ antibody
    • H3FK antibody
    • H3FL antibody
    • HIST1H3A antibody
    • HIST1H3B antibody
    • HIST1H3C antibody
    • HIST1H3D antibody
    • HIST1H3E antibody
    • HIST1H3F antibody
    • HIST1H3G antibody
    • HIST1H3H antibody
    • HIST1H3I antibody
    • HIST1H3J antibody
    • HIST3H3 antibody
    • Histone 1, H3a antibody
    • Histone 1, H3b antibody
    • Histone 1, H3c antibody
    • Histone 1, H3d antibody
    • Histone 1, H3e antibody
    • Histone 1, H3f antibody
    • Histone 1, H3g antibody
    • Histone 1, H3h antibody
    • Histone 1, H3i antibody
    • Histone 1, H3j antibody
    • Histone cluster 1, H3a antibody
    • Histone cluster 1, H3b antibody
    • Histone cluster 1, H3c antibody
    • Histone cluster 1, H3d antibody
    • Histone cluster 1, H3e antibody
    • Histone cluster 1, H3f antibody
    • Histone cluster 1, H3g antibody
    • Histone cluster 1, H3i antibody
    • Histone cluster 1, H3j antibody
    • Histone H 3 antibody
    • Histone H3.1 antibody
    • Histone H3.2 antibody
    • Histone H3/a antibody
    • Histone H3/b antibody
    • Histone H3/c antibody
    • Histone H3/d antibody
    • Histone H3/f antibody
    • Histone H3/h antibody
    • Histone H3/i antibody
    • Histone H3/j antibody
    • Histone H3/k antibody
    • Histone H3/l antibody
    • Histone H3/m antibody
    • Histone H3/o antibody
    see all

Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade images

  • Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The  ChIP was performed with 25µg of chromatin, 2µg of  ab8895 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified on the GAPDH and ALDOA (active) and MYO-D (inactive) promoters and over the y-Actin gene (active). Schematic diagram of the y-Actin gene is shown on the top of the figure. Black boxes represent exons and thin lines represent introns. PCR products are depicted as bars under the gene.

  • A: Enlarged region showing the distribution of the H3K4me1 labelling around and within the EC cavities of a murine rod photoreceptor.
    B: Schematic representation of the labelling for the H3K4me1 mark. The bar represents 260 nm in A and 1 µm in B.
  • ab8895 staining Histone H3 (mono methyl K4) in HeLa cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab8895 at 1µg/ml and ab7291 (anti beta Tubulin) at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat anti-rabbit AlexaFluor®488 secondary (ab150081) at 2 μg/ml (shown in green) and a goat anti-mouse AlexaFluor®594 secondary (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.

    Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.

  • IHC image of ab8895 staining Histone H3 (mono methyl K4) in human colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab8895, 0.5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).

     

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • All lanes : Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895) at 1/500 dilution

    Lane 1 : Calf thymus histone lysate
    Lane 2 : Calf thymus histone lysate with Human Histone H3 (mono methyl K4) peptide (ab1340) at 1 µg/ml
    Lane 3 : Calf thymus histone lysate with Human Histone H3 (di methyl K4) peptide (ab7768) at 1 µg/ml
    Lane 4 : Calf thymus histone lysate with Human Histone H3 (tri methyl K4) peptide (ab1342) at 1 µg/ml
    Lane 5 : Calf thymus histone lysate with Human Histone H3 (mono methyl K9) peptide (ab1771) at 1 µg/ml
    Lane 6 : Calf thymus histone lysate with Human Histone H3 (mono methyl K27) peptide (ab1780) at 1 µg/ml
    Lane 7 : Calf thymus histone lysate with Human Histone H3 (unmodified ) peptide (ab2903) at 1 µg/ml

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution

    Performed under reducing conditions.

    Predicted band size : 15 kDa


    Exposure time : 2 minutes

    ab8895 is specific for mono-methylated Lysine 4 of histone H3 and does not recognize di- or tri-methyl Lysine 4 nor methylation at Lysine 9. This is shown in lane 2 where the activity of the antibody is specifically blocked by the addition of the immunizing peptide (ab1340).

  • Indian muntjac fibroblast cells -  interphase (top left) and prophase (top right and below), stained with:

    Mono Methyl K4 antibody, ab8895, (green)
    DAPI: red, top left and right; blue, below
    Phospho Ser 10 antibody: red (below) and blue (top right)

    The perinuclear and perinucleolar heterochromatin domains do not contain Mono Methyl K4.  The Mono Methyl K4, rather, is distributed as small nuclear foci primarily found between DAPI-intense regions of the nucleus. 

     

  • ab8895 used in Dot Blot at a 1/1000 dilution, 18 hours at 4°C.
    The dot blot was done using 0.1ug of peptide and screened multiple histone tail modifications to ensure no cross reactivity.

    See Abreview

  • ab8895 staining mouse embryonic stem cells by flow cytometry. The ES cell colonies were trypsinized and the cells permeabilized before being stained with ab8895 (0.25ug/1.5 x 105 cells). A PE conjugated goat anti-rabbit antibody was used as the secondary.

    See Abreview

References for Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (ab8895)

This product has been referenced in:
  • Pradeepa MM  et al. Histone H3 globular domain acetylation identifies a new class of enhancers. Nat Genet N/A:N/A (2016). Human . Read more (PubMed: 27089178) »
  • Zhang J  et al. Disruption of KMT2D perturbs germinal center B cell development and promotes lymphomagenesis. Nat Med N/A:N/A (2015). WB, ChIP ; Mouse . Read more (PubMed: 26366712) »

See all 187 Publications for this product

Product Wall

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Trachemys scripta elegans Tissue lysate - whole (Liver)
Gel Running Conditions Reduced Denaturing (Tris-glycine, 15%)
Loading amount 25 µg
Specification Liver
Blocking step High Molecular Weight PVA as blocking agent for 1 minute(s) · Concentration: 1000µg/mL · Temperature: RT°C
Username

Mr. Liam Hawkins

Verified customer

Submitted Jan 20 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Western blot
Sample Ictidomys tridecemlineatus Tissue lysate - other (Muscle)
Gel Running Conditions Reduced Denaturing (Tris-tricine, 15%)
Loading amount 35 µg
Specification Muscle
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: RT°C
Username

Mr. Liam Hawkins

Verified customer

Submitted Jan 15 2016

Application Western blot
Sample Rana sylvatica Tissue lysate - whole (Liver)
Gel Running Conditions Reduced Denaturing (15% Tris-glycine)
Loading amount 40 µg
Specification Liver
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: RT°C
Username

Mr. Liam Hawkins

Verified customer

Submitted Jan 15 2016

Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (testis)
Permeabilization Yes - 0.5% Triton
Specification testis
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 08 2015

Abcam has not validated the combination of species/application used in this Abreview.
Application ChIP
Sample Zebrafish Cell lysate - nuclear (whole embryo shield stage, 6 hours postferttilizat)
Negative control We used PCR for zgc15 and igsf genomic regions (see additional information for primer sequences) as a reference control. No H3K4 mono- and di- methylation was expected in these regions based on previously published microarray data
Specification whole embryo shield stage, 6 hours postferttilizat
Detection step Real-time PCR
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde in PBST
Positive control ChIP sample was not enriched over input control in hoxb3a or in tiparp genomic regions (see additional information for primer sequences). 10-20x enrichment in H3K4 mono- and di- methylation was expected in these regions based on previously published microarray data.
Username

Mrs. Daria Onichtchouk

Verified customer

Submitted Apr 29 2015

Application Immunohistochemistry (Frozen sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C
Sample Human Tissue sections (human colon)
Specification human colon
Permeabilization No
Fixative Acetone
Username

Abcam user community

Verified customer

Submitted Dec 01 2014

Application Western blot
Loading amount 25 µg
Gel Running Conditions Non-reduced Denaturing (4-12)
Sample Xenopus tropicalis Tissue lysate - whole (WHOLE EMBRYO)
Specification WHOLE EMBRYO
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Nov 03 2014

Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Na-citrate pH6
Sample Mouse Tissue sections (Liver, P5)
Specification Liver, P5
Permeabilization Yes - Triton 0,05%
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 05 2014

As both lots are at 1mg/ml concentration, they are BSA free. We add this only to lots below 1mg/ml as a stabilizing agent. Unfortunately, we are unable to do bespoke BSA free orders for such small volumes.

Here is our in-house ChIP protocol:

Scope This standard operation procedure is written to provide instructions for the method of Chromatin immunoprecipitation using fixed chromatin.
Safety considerations Safety gloves, a laboratory coat,...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Marmoset (common) Tissue sections (adult marmoset testis)
Specification adult marmoset testis
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Dako Antigen retrieval solution
Permeabilization No
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Zachary Yu-Ching Lin

Verified customer

Submitted Sep 27 2012

For ChIP in general, the Abcam lab always does the IP step overnight (16 hours) at 4C with rotation. For the beads, they prepare a 50:50 mix of protein A/G Sepharose beads (GE Healthcare UK LTD). They typically prepare a new bead slurry every month or ...

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At this time we do not have a PE-conjugated version of ab8895.
As an alternative, you may be interested in our Easylink conjugation kits. These simple kits allow one to conjgate a variety of fluors, enzymes, etc to any antibody. We have PE conjuga...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Plasmodium falciparum Cell lysate - nuclear (Acid extracted histones from trophozoites)
Loading amount 1e+007 cells
Specification Acid extracted histones from trophozoites
Gel Running Conditions Reduced Denaturing (4-12% Bis-Tris)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jul 11 2012

The concentration of ab8895 varies by the specific lot that you received.

After over 13,000 individual ELISA experiments, we have determined that storing our antibodies at various temperatures of up to 45°C for 1 week does not impact their activity. Our Abpromise to you is that, should our product not work as stated on t...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (MCF-7)
Specification MCF-7
Fixative Formaldehyde
Permeabilization Yes - Triton-X-100
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Abcam user community

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Submitted May 14 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (SAOS2 HUMAN OSTEOSARCOMA)
Specification SAOS2 HUMAN OSTEOSARCOMA
Fixative Paraformaldehyde
Permeabilization Yes - PBS-TRITON 0.25%
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted May 04 2012

Currently, we do not offer full length methylated H3K4 proteins. We do have an EpiSeeker Histone Extraction Kit (ab113476) which provides a simple and selective method for extracting histone proteins in just 1 hour. When used in conjunction with our pe...

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I can confirm that we sell the relevant modified peptides, however currently we do not manufacture ready-to-use dot blots or peptide arrays. For further information on ChIP techniques, please visit our Protocol sections: http://www.abcam.com/index.htm...

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We have 2 antibodies that should be suitable for your research work :
- ab4729, Anti-Histone H3 (acetyl K27) antibody - ChIP Grade, www.abcam.com/ab4729
- ab8895, Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade, www.abcam.com/ab8895 ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP on chip
Sample Mouse Cell lysate - other (Histone peptide array)
Specification Histone peptide array
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Detection step Microarray
Username

Dr. Michael Chopin

Verified customer

Submitted Sep 22 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Saccharomyces cerevisiae Cell lysate - whole cell (S288c)
Loading amount 25 µg
Specification S288c
Gel Running Conditions Reduced Denaturing (15% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Mr. Harsh Ja

Verified customer

Submitted Aug 02 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Saccharomyces cerevisiae Cell lysate - nuclear (S288c)
Specification S288c
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Fromaldehyde
Detection step Real-time PCR
Username

Mr. Harsh Ja

Verified customer

Submitted Aug 02 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Saccharomyces cerevisiae Cell lysate - nuclear (BY4147 background)
Total protein in input 100000 cells
Specification BY4147 background
Immuno-precipitation step Protein G
Username

Abcam user community

Verified customer

Submitted Jul 27 2011

I have heard back from the ChIP expert in the lab, and I was told the following: "Unfortunately, I cannot comment on cell line specific gene enrichment BUT we have noticed in the past that the MYO-D loci is the least stable in Jurkat’s. I cannot com...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Saccharomyces cerevisiae Cell lysate - nuclear (BY4741)
Loading amount 10 µg
Specification BY4741
Gel Running Conditions Reduced Denaturing (15% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

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Submitted Jun 01 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP on chip
Sample Saccharomyces cerevisiae Cell lysate - nuclear (BY4741)
Specification BY4741
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde
Detection step Microarray
Positive control WT cells and set1
Negative control no antibody control
Username

Abcam user community

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Submitted May 31 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry
Sample Saccharomyces cerevisiae Cell (W303 cells)
Specification W303 cells
Fixative Paraformaldehyde
Permeabilization Yes - .25% Triton X-100 in PBS
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 25°C
Username

Abcam user community

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Submitted May 31 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Saccharomyces cerevisiae Cell lysate - nuclear (BY4741 cells)
Specification BY4741 cells
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 20 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde
Detection step Semiquantitative PCR
Username

Abcam user community

Verified customer

Submitted May 26 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Mouse Cell lysate - nuclear (MEF)
Specification MEF
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldehyde
Detection step Other
Username

Abcam user community

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Submitted Nov 22 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - nuclear (K562 - 1 million cells per IP)
Specification K562 - 1 million cells per IP
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldehyde
Detection step Other
Username

Abcam user community

Verified customer

Submitted Nov 01 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Saccharomyces cerevisiae Cell lysate - whole cell (cell lysate and purified nucleosomes)
Loading amount 40 µg
Specification cell lysate and purified nucleosomes
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Username

Dr. Joseph Reese

Verified customer

Submitted Feb 01 2010

Unfortunately, we cannot provide sequences of primers since they are intellectual property. But now we are providing each primer pairs and probes as follows; ab85378 Human GAPDH exon 1 ChIP Primer and Probe Set, ab85773 Human Aldolase intron 2 ChIP Pri...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - whole cell (U2OS)
Specification U2OS
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde 1%
Detection step Taqman
Positive control Histone H3 (ab1791)
Negative control protein A beads
Username

Abcam user community

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Submitted Dec 03 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.2% Triton in PBS
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
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Submitted Jun 18 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Saccharomyces cerevisiae Cell lysate - whole cell (N/A)
Loading amount 100000 cells
Specification N/A
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Oct 23 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Mouse Cell (Embryonic stem cells)
Specification Embryonic stem cells
Preparation Cell harvesting/tissue preparation method: ES cell colonies were trypsinized and strained through cell strainer
Sample buffer: Cells were kept in FACS buffer or PBS
Fixation Commercial Fixation concentrate
Permeabilization Yes - Commercial Permeabilization Buffer
Gating Strategy All living cells were gated
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Ms. Nadine Obier

Verified customer

Submitted Aug 21 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Saccharomyces cerevisiae Cell lysate - whole cell (Whole cell extract)
Loading amount 40 µg
Specification Whole cell extract
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
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Submitted Jun 19 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (zygotes)
Specification zygotes
Fixative Paraformaldehyde
Blocking step BSA as blocking agent for 10 hour(s) and 0 minute(s) · Concentration: 1%
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Dr. Konstantin Lepikhov

Verified customer

Submitted Apr 04 2007

All antibodies that are batch tested here at Abcam have the primers that were employed during the testing published as a separate page which can be accessed as a clickable link on the antibody datasheet. For example on the antibody datasheet for ab8895...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (thymus)
Loading amount 25000 cells
Specification thymus
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
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Submitted Oct 31 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
Application ChIP
Sample Human Cell lysate - whole cell (HeLa)
Specification HeLa
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Detection step Other
Positive control TAF p250, sc-735
Negative control no antibody ChIP-chip
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Miss. Rhona Stuart

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Submitted Jul 07 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Not applicable Cell lysate - whole cell (whole cell lysate - C. albicans)
Loading amount 2 µg
Specification whole cell lysate - C. albicans
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5%
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Dr. Suresh Babu Raman

Verified customer

Submitted Mar 24 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Saccharomyces cerevisiae Cell lysate - whole cell
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
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Abcam user community

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Submitted Jan 27 2006

I am pleased to confirm that we have tested our antibodies ab8895 and ab7766 with a non-modified blocking peptide and the antibodies performed very well, we therefore believe they are specific to modified histones only.

You are correct in your statement that the antibodies have not been selected against other methylated forms. However, we can state that the purified antibody is specific for the mono or di, as we test the antibody with blocking peptides to the other me...

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Ab8895 is specific for mono-methyl K4 and Ab7766 is specific for di-methyl K4. This is achieved by affinity purification with the immunizing peptide.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"