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Anti-Histone H3 (methylated) antibody [mAbcam 6002] - ChIP Grade (ab6002)

Abreviews (35)Q&A (69)Specific References (58)

Overview

  • Product nameAnti-Histone H3 (methylated) antibody [mAbcam 6002] - ChIP GradeSee all Histone H3 (methylated) primary antibodies ...
  • Description
    Mouse monoclonal [mAbcam 6002] to Histone H3 (methylated) - ChIP Grade
  • SpecificityThis antibody is specific for histone H3 tri-methylated at K27. The sequence is found in all mammals and a wide range of species, including D. melanogaster, Arabidopsis, Chicken and Xenopus. The antibody will react with any of the above species where the modification is present. Reactivity was confirmed using Cow samples. Reactivity is not certain in S. pombe and S. cerevisiae as the equivalent protein sequence differs slightly from the species listed above. A customer reported that the activity against histone H3 in Zea mays (maize) is not good. The antibody is blocked in western blotting by tri methyl K27 peptide and slightly by di methyl K27 peptide. It is not blocked by mono methyl K4, di methyl K4, tri methyl K4, mono methyl K9, di methyl K9, tri methyl K9, mono methyl K27 or unmodified K27 peptides. (See blot below).
  • Tested applicationsChIP/Chip, ChIP, IHC-P, Flow Cyt, WB, ICC, ICC/IF more details
  • Species reactivity
    Reacts with: Mouse, Cow, Human, Arabidopsis thaliana, Fruit fly (Drosophila melanogaster), Plants, Zebrafish, Rhesus monkey
    Predicted to work with: Rat, Rabbit, Chicken, Xenopus laevis, Chinese Hamster
  • Immunogen

    Synthetic peptide derived from residues 1 - 100 of Human Histone H3, tri methylated at K27.

    Read Abcam's proprietary immunogen policy

    . Hybridomas were prepared and the resulting clones were positively screened by ELISA against the immunising peptide. Clones were negatively screened against both the corresponding unmodified peptide and also against a peptide corresponding to tri methylated K9 of Histone H3.

  • General notesNOT SUITABLE for blocking with milk. Block in 5% BSA for 1 hour. Our labs have investigated the blocking conditions for this antibody and found that milk significantly decreases the signal and is therefore not a suitable blocking agent for this antibody (see Western Blot image).

Properties

Applications

Our Abpromise guarantee covers the use of ab6002 in the following tested applications.

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

Application Notes
ChIP/Chip ChIP/Chip: Use at an assay dependent dilution.
ChIP ChIP: Use 5-10 µg for 25 µg of chromatin.
IHC-P IHC-P: Use at an assay dependent dilution.
Flow Cyt Flow Cyt: Use at an assay dependent dilution.
WB WB: Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Histone H3 peptide - tri methyl K27 (ab1782). Block with BSA. The signal is very weak with milk blocking.
ICC ICC: Use at an assay dependent dilution.
ICC/IF ICC/IF: Use a concentration of 5 µg/ml.

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.

    • Target information above from: UniProt accession P68431 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
    see all
  • Alternative names
      FLJ92264 antibodyH3 histone antibodyH3 histone antibody
      H3 histone family, member A antibodyH3/a antibodyH3/b antibodyH3/c antibodyH3/d antibodyh3/f antibodyH3/h antibodyH3/i antibodyH3/j antibodyH3/k antibodyH3/l antibodyH31_HUMAN antibodyH3F1K antibodyH3F3 antibodyH3F3 antibodyH3FA antibodyH3FB antibodyH3FC antibodyH3FD antibodyH3FF antibodyH3FH antibodyH3FI antibodyH3FJ antibodyH3FK antibodyH3FL antibodyHIST1H3A antibodyHIST1H3B antibodyHIST1H3C antibodyHIST1H3D antibodyHIST1H3E antibodyHIST1H3F antibodyHIST1H3G antibodyHIST1H3H antibodyHIST1H3I antibodyHIST1H3J antibodyHIST3H3 antibodyHIST3H3 antibodyHistone 1, H3a antibodyHistone cluster 1, H3a antibodyHistone cluster 1, H3b antibodyHistone cluster 1, H3c antibodyHistone cluster 1, H3d antibodyHistone cluster 1, H3e antibodyHistone cluster 1, H3f antibodyHistone cluster 1, H3g antibodyHistone cluster 1, H3i antibodyHistone cluster 1, H3j antibodyHistone H 3 antibodyHistone H3.1 antibodyHistone H3.1 antibodyHistone H3/a antibodyHistone H3/b antibodyHistone H3/c antibodyHistone H3/d antibodyHistone H3/f antibodyHistone H3/h antibodyHistone H3/i antibodyHistone H3/j antibodyHistone H3/k antibodyHistone H3/l antibody
    see all

Anti-Histone H3 (methylated) antibody [mAbcam 6002] - ChIP Grade images

  • ChIP - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    ChIP - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The  ChIP was performed with 25µg of chromatin, 5µg of  ab6002 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach for active and inactive loci, Sybr green approach for heterochromatic loci). Primers and probes are located in the first kb of the transcribed region.
        
  • Western blot - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    Western blot - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    All lanes : Anti-Histone H3 (methylated) antibody [mAbcam 6002] - ChIP Grade (ab6002) at 1 µg/ml

    Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate
    Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide (ab17163) at 0.5 µg/ml
    Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - mono methyl K4 (ab1340) at 0.5 µg/ml
    Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - di methyl K4 (ab7768) at 0.5 µg/ml
    Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - tri methyl K4 (ab1342) at 0.5 µg/ml
    Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - mono methyl K9 (ab1771) at 0.5 µg/ml
    Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - di methyl K9 (ab1772) at 0.5 µg/ml
    Lane 8 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - tri methyl K9 (ab1773) at 0.5 µg/ml
    Lane 9 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - mono methyl K27 (ab1780) at 0.5 µg/ml
    Lane 10 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - di methyl K27 (ab1781) at 0.5 µg/ml
    Lane 11 : Calf Thymus Histone Preparation Nuclear Lysate with Histone H3 peptide - tri methyl K27 (ab1782) at 0.5 µg/ml

    Lysates/proteins at 0.5 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 15 kDa
    Observed band size : 17 kDa (why is the actual band size different from the predicted?)


    Exposure time : 3 minutes
  • Western blot - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    Western blot - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    All lanes : Anti-Histone H3 (methylated) antibody [mAbcam 6002] - ChIP Grade (ab6002) at 1 µg/ml

    Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg with BSA BLOCK
    Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate at 0.5 µg with MILK BLOCK
    Lane 3 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg with BSA BLOCK
    Lane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg with BSA BLOCK
    Lane 5 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg with MILK BLOCK
    Lane 6 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg with MILK BLOCK

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 15 kDa
    Observed band size : 17 kDa (why is the actual band size different from the predicted?)


    Exposure time : 12 minutes
  • Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)Luke Hughes-Davies and Rhiannon Jade, Gurdon Institute, Cambridge, UK

    Immunofluorescent imaging of human cells (U2OS) with ab6002 reveals broadly dispersed interphase nuclear staining corresponding to trimethylation of K27, with multiple foci of brighter staining, exactly agreeing with published studies of K27-trimethyl IF [See figure 3 of Peters et al Mol Cell 12(6):1577-1589 (2003)] .  The lack of nucleolar or cytoplasmic staining background confirms the high specificity of the antibody in this application.   

    IF was performed with a standard paraformaldehyde technique (fixed in PBS buffered PFH 4% for 5 minutes, permeabilised with 0.5% triton-PBS for 5 minutes, blocked with 5% milk / 0.2% tween for one hour.  Primary antibody used at 1/200 in 5% milk / 0.2% TWEEN for one hour, secondary antibody Alexa 488 for 30 minutes.  All blocking and incubation steps carried out at 37 degrees C. 

  • Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)

    Figure showing the nuclear distribution of H3 (tri-methyl K27) antibody, ab6002 in a) a 46 chromosome, XX cell line, and b) a 49 chromosome, XXXXX cell line.

    The location of facultative heterochromatin at the inactive X chromosome is indicated by white arrow heads.

  • Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)Kirk Mcmanus
    Interphase 10T1/2 mouse fibroblasts were paraformaldehyde fixed (4%), immunofluorescently labeled with anti-trimethyl K27 antibody (ab6002) and counterstained with DAPI.  The merge image presents the DAPI and ab6002 channels as red and green, respectively.  The scale bar represents 3µm.
  • ELISA - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    ELISA - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)

    ELISA using ab6002 at antibody concentrations between 1/500 and 1/8000.

    The red line indicates binding to the tri methyl K27 peptide (ab1782). Binding to the following peptides was not seen:
    Unmodified K27 (ab2623),
    mono methyl K27 (ab1780),
    di methyl K27 (ab1781),
    mono methyl K9 (ab1771),
    di methyl K9 (ab1772),
    tri methyl K9 (ab1773),
    mono methyl K4 (ab1340),
    di methyl K4 (ab7768),
    tri methyl K4 (ab1342).

    This indicates the specificity of ab6002 for tri methyl K27 of Histone H3.

  • Flow Cytometry - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    Flow Cytometry - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)This image is courtesy of an Abreview submitted by Prof Albrecht Müller
    ab6002 staining mouse embyronic stem cells by flow cytometry. The ES cell colonies were trypsinized and permeabilized prior to blocking and staining with the antibody (1ug/1.5 x 10 5 cells. A Cy3 conjugated anti-mouse antibody was used as the secondary.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    Immunocytochemistry/ Immunofluorescence - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    ICC/IF image of ab6002 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6002, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 5µg/ml, and in 100% methanol fixed (5 min) HeLa, Hek293, HepG2 and MCF7 cells at 5µg/ml.
  • ChIP - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    ChIP - Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)This image is a courtesy of Steve Bilodeau

    Chromatin was prepared from nuclear lysate of the mouse embryonic stem cells. The cross-linking (X-ChiP) technique was used, crosslinking was performed for 15 minutes in 1% formaldehyde. The primary antibody was diluted to 0.0133µg/µg chromatin and incubated in ChIP Sonication Buffer with the sample for 24 hours at 4°C. The immunoprecipitated DNA was quantified by real time PCR.

    Cdx2: PCR primers situated in the promoter regions of Caudal type homeobox transcription factor 2
    Nef3: PCR primers situated in the promoter regions of neurofilament 3

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002)
    ICC/IF image of ab6002 stained HepG2 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab6002, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-Histone H3 (methylated) antibody [mAbcam 6002] - ChIP Grade (ab6002)

This product has been referenced in:
  • Jiao B  et al. 8-CPT-cAMP/all-trans retinoic acid targets t(11;17) acute promyelocytic leukemia through enhanced cell differentiation and PLZF/RARa degradation. Proc Natl Acad Sci U S A 110:3495-500 (2013). ChIP ; Mouse . Read more (PubMed: 23382200) »
  • Mysliwiec MR  et al. Jarid2 (Jumonji, AT Rich Interactive Domain 2) Regulates NOTCH1 Expression via Histone Modification in the Developing Heart. J Biol Chem 287:1235-41 (2012). ChIP ; Mouse . Read more (PubMed: 22110129) »

See all 58 Publications for this product

Publishing research using ab6002 ? Please let us know so that we can cite the reference in this datasheet

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Displaying 1 - 10 of 104 results for Abreviews and Q&A

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Votes: Highest First

I will be using the H3K27me3 antibody (ab6002) with the specification that it doesn’t work with blocking with milk and BSA should be used instead. We prepare our primary dilutions in 1% milk in PBS-T, is this antibody suitable to dilute in milk?



We do not recommend to mix blocking agents in general.

In the case of ab6002 it is especially important to use BSA only. Using milk in blocking or antibody solutions will lead to a weak signal.

Therefore I recommend to add BS...

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Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (methylated) antibody [mAbcam 6002] - ChIP Grade

 Average
Abreviews
Application Immunocytochemistry/ Immunofluorescence
Sample Cyanidioschyzon merolae Cell (Synchronous culture using light/dark cycle)
Specification Synchronous culture using light/dark cycle
Fixative Paraformaldehyde
Permeabilization No
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 37°C
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Submitted Feb 12 2013

Western blot abreview for Anti-Histone H3 (methylated) antibody [mAbcam 6002] - ChIP Grade

 Good
Abreviews
Application Western blot
Sample Human Cell lysate - whole cell (Cell lines)
Loading amount 5 µg
Specification Cell lines
Gel Running Conditions Reduced Denaturing
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Feb 01 2013

Application: IHC-P
Sample: Human prostate tissue
AR: EDTA
Primary: 1/100, 1/200. O/N at 4oC
Detection: Lyca powervision plus 30', TAB 20' : works with other antibodies
This antibody has worked multiple times in the past with consistent...

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We recently spoke regarding ab6002, specifically the lack of positive signal during your optimization attempts using a protocol that worked for you previously. As we discussed, I am sending you a free of charge replacement vial of the same antibody u...

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I'm doing a dot blot peptide blocking study and I'm seeing reactivity in both the non-methylated and trimethylated peptides.

Thank you for contacting us.

I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our...

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ChIP abreview for Anti-Histone H3 (methylated) antibody [mAbcam 6002] - ChIP Grade

 Good
Abreviews
Application ChIP
Sample Human Cell lysate - nuclear (AML)
Specification AML
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Detection step Real-time PCR
Positive control PAX5
Negative control PU.1
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Verified customer

Submitted Nov 19 2012

Dear Sir/Madam,
I am looking to purchase some antibodies from your company, and was wondering if there is any way of getting a trial size, or discount on antibodies that have not yet being tested on certain species? I work on chicken cells and testin...

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Thank you for contacting us and for your interest in our products.

I am very sorry to hear that ab6002 didn't work in your samples. Have you already contacted Scientific support about this to ask for protocol tips?

Unfortunately, bec...

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ChIP abreview for Anti-Histone H3 (methylated) antibody [mAbcam 6002] - ChIP Grade

 Excellent
Abreviews
Application ChIP
Sample Mouse Cell lysate - other (Mouse ES cells)
Specification Mouse ES cells
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 12 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% formaldehyde in culture med
Detection step Real-time PCR
Positive control En1 promoter (Engrailed homologue 1, well characterized polycomb target in ES cells)
Negative control Oct4 promoter
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Submitted Nov 05 2012

ChIP abreview for Anti-Histone H3 (methylated) antibody [mAbcam 6002] - ChIP Grade

 Excellent
Abreviews
Application ChIP
Sample Human Cell lysate - whole cell (Human medulloblastoma cells)
Specification Human medulloblastoma cells
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 Semiquantitative PCR
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Submitted Oct 18 2012

Dear Sir/Madam,

We use this antibody for westerns. Upon receiving the antibody, however, we intended to use it for ChIP, so we aliquoted it to rather high volumes and stored it at -80C. Now that I need it for westerns, I have thawed a batch, bu...

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Thank you for your enquiry.

I can confirm we would recommend to avoid repeat freeze-thawing of an antibody.

I would recommend if you are going to use the antibody quite soon in the next few weeks to keep it at 4oC.

If you ...

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