Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

Overview

  • Product name
    Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade
    See all Histone H3 primary antibodies
  • Description
    Rabbit polyclonal to Histone H3 - Nuclear Loading Control and ChIP Grade
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-Fr, CHIPseq, Dot blot, Flow Cyt, IHC-P, Electron Microscopy, ICC/IF, ChIP, IP, WB, ChIP/Chip, IHC - Wholemount, ICCmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Dog, Human, Saccharomyces cerevisiae, Xenopus laevis, Arabidopsis thaliana, Caenorhabditis elegans, Drosophila melanogaster, Ferret, Indian muntjac, Schizosaccharomyces pombe, Zebrafish, Silk worm, Dictyostelium discoideum, Rainbow trout, Trypanosoma cruzi, Neurospora crassa, Toxoplasma gondii, Rice, Schistosoma mansoni, Candida albicans, Cyanidioschyzon merolae
    Predicted to work with: a wide range of other species, Mammals
  • Immunogen

    Synthetic peptide corresponding to Human Histone H3 aa 100 to the C-terminus conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: P68431
    (Peptide available as ab12149)

  • Positive control
    • WB: HeLa, Drosophila embryo nuclear extract, NIH/3T3, S.cerevisiae (Y190) and S.pombe whole cell lysates. ICC/IF: Methanol fixed HeLa cells. ChIP: Chromatin from HeLa cells.
  • General notes

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

    We recommend Goat Anti-Rabbit IgG H&L (HRP) (ab6721) secondary antibody.

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped 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 buffer
    pH: 7.40
    Preservative: 0.01% 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...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab1791 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
IHC-Fr 1/200.
CHIPseq Use at an assay dependent concentration.
Dot blot 1/10000.
Flow Cyt Use at an assay dependent concentration.

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

IHC-P 1/100 - 1/400. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Electron Microscopy 1/50. Customer feedback
ICC/IF Use a concentration of 1 µg/ml.

Methanol fixed cells.

ChIP Use 2µg for 106 cells.
IP Use a concentration of 5 µg/ml.
WB 1/1000 - 1/5000. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).Can be blocked with Human Histone H3 peptide (ab12149).

We recommend Goat Anti-Rabbit IgG H&L (HRP) (ab6721) secondary antibody.

ChIP/Chip Use at an assay dependent concentration.
IHC - Wholemount Use at an assay dependent concentration. PubMed: 22219645
ICC 1/100 - 1/500.

Target

  • Function
    Core 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 similarities
    Belongs to the histone H3 family.
  • Developmental stage
    Expressed 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 localization
    Nucleus. Chromosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • H3 histone family member E pseudogene antibody
    • H3 histone family, member A antibody
    • H3/A antibody
    • H31_HUMAN antibody
    • H3F3 antibody
    • H3FA 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 cluster 1, H3a antibody
    • Histone H3 3 pseudogene antibody
    • Histone H3.1 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
    see all

Images

  • H3 oxidation in major satellite region was determined by sequential ChIP (re-ChIP) in control and SNA1 KO MEFs. The lysate was incubated with biotin-hydrazide (an activated biotin that reacts with oxidized H3) before re-ChIP. Extracts were sequentially immunoprecipitated with anti-H3 and streptavidin beads. DNA binding was quantified by qPCR. Data were normalized to the total amount of H3 immunoprecipitated and to the input and expressed as fold enrichment over the data obtained when an irrelevant IgG was used. 

    See Abreview

  • Chromatin from Xenopus laevis oocytes was prepared according to the Abcam X-ChIP protocol.

    Oocytes were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25 mg of chromatin, 3 mg of ab7834 (anti-H3, light blue) and 3 µg of ab1791 (anti-H3, dark blue), and 20 ml of Protein A/G sepharose beads. A non-specific antibody was used as a control (yellow).

    The immunoprecipitated DNA was quantified by real time PCR (Taqman approach).

  • All lanes : Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791) at 1/1000 dilution

    Lane 1 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 4 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with Human Histone H3 peptide (ab12149) at 1 µg/ml
    Lane 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Human Histone H3 peptide (ab12149) at 1 µg/ml
    Lane 6 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate with Human Histone H3 peptide (ab12149) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

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

    Developed using the ECL technique.

    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: 10 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab1791 overnight at 4°C.

    Goat Anti-Rabbit IgG H&L (HRP) (ab6721) secondary antibody was used for detection.

    Antibody binding was visualised using ECL development solution ab133406.

  • All lanes : Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : NIH 3T3 whole cell lysate (ab7179)
    Lane 3 : Drosophila embryo nuclear extract (from melanogaster embryos 0-12Hr)
    Lane 4 : S.cerevisiae (Y190) Whole Cell Lysate
    Lane 5 : S.pombe Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Rabbit 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?



    ab1791 is tested in western blot on a range of species.  We recommend loading higher amounts of protein (20-30ug) to increase the signal in yeast lysates
  • Chromatin was prepared from HeLa (Human epithelial cell line from cervix adenocarcinoma) cells according to the Abcam X-ChIP protocol.

    Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 25 µg of chromatin, 2 µg of ab1791 (blue), and 20 µl 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.

  • Rabbit polyclonal to Histone H3 (ab1791) at 1/5000 on S. cerevisiae whole cell lysate (40 ug per lane).

    Protein resolved on 15% SDS-PAGE gel. After transfer to PVDF membrane, blots were blocked in 1X PBS, 0.1% Tween-20, and 5% milk. ab1791 was diluted in 5 ml blocking buffer at 1/5000.  Blots plus primary antibodies were either incubated overnight at 4°C or at RT for 2 hours. Blots were washed 6X for 10 minutes each in PBS with 0.1% Tween-20 before addition of secondary antibodies. Secondary antibodies were diluted 1/2,000 in blocking buffer and incubated with blots for 2 hours at RT. Secondary blots were washed 4X for 10 minutes each in PBS with 0.1% Tween-20 and 2X for 10 minutes each in PBS.

  • ab1791 staining Histone H3 in HeLa (Human epithelial cell line from cervix adenocarcinoma) by ICC/IF (Immunocytochemistry/immunofluorescence).

    Cells were fixed with methanol and blocked with 0.2% fish scale gelatin for 1 hour at 25°C. Samples were incubated with the primary antibody (1/300 in PBS + 0.2% gelatin) for 20 minutes at 25°C. An Alexa Fluor® 488-conjugated donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

    Green - Histone H3.
    Blue - DAPI.
    Red - Tubulin.

    See Abreview

  • Paraffin-embedded rat brain tissue stained for Histone H3 using ab1791 at 1/8000 dilution in immunohistochemical analysis.

    See Abreview

  • The ChIP was performed with chromatin from mouse gut cell lysate and ab1791 at 1/250 dilution.

    Negative control: No antibody was used (right bar).

    The immunoprecipitated DNA was quantified by real time PCR.

     

    See Abreview

  • ab1796 staining Histone H3 in mouse liver tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

    Tissue was fixed with paraformaldehyde, permeabilized with 0.05% Triton X-100 in PBS for 30 minutes and blocked with 5% BSA for 1 hour; antigen retrieval was by heat mediation in sodium citrate pH 6. Samples were incubated with the primary antibody (1/500 in blocking buffer) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.

    See Abreview

  • Histone H3 - ChIP Grade was immunoprecipitated using 0.5mg HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell extract, 5 µg of Rabbit polyclonal to and 50 µl of protein G magnetic beads (+). No antibody was added to the control (-).

    The antibody was incubated under agitation with Protein G beads for 10 minutes, HeLa whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10 minutes under agitation.

    Proteins were eluted by addition of 40 µl SDS loading buffer and incubated for 10 minutes at 70°C; 10 µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab1791.

    Secondary Antibody: Mouse anti-rabbit HRP light chain (HRP) (ab99697).

    Band: 15kDa; Histone H3 - ChIP Grade

  • ab1791 staining Histone H3 in human infantile fibromatosis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

    Tissue was fixed with formaldehyde and blocked with 1% FBS/BSA for 3 hours at room temperature; antigen retrieval was by heat mediation in Tris pH 9. Samples were incubated with primary antibody (1/100 in TBS + 1% BSA + 1% FBS) for 16 hours. An un-diluted HRP-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

  • ab1791 staining Histone H3 (red) in rat brain tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

    Tissue was fixed with formaldehyde, permeabilized with 0.1% TBS-TritonX and blocked with 10% serum for 1 hour at 25°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with the primary antibody (1/500 in 10% normal goat serum) for 24 hours at 24°C. An Alexa Fluor® 594-conjugated goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

    Green - Nucleus staining.
    Red - Histone H3 staining.

  • All lanes : Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791) at 1/1000 dilution

    All lanes : Mouse skeletal muscle mitochondrial fraction

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG at 1/4000 dilution

    Developed using the ECL technique.

    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: 7 minutes


    Blocked with 3% milk for 1 hour at 25°C.

    Incubated with the primary antibody for 16 hours at 4°C in 3% milk in TBS-tween.

    See Abreview

  •  ab1791 staining mouse embryonic stem cells by flow cytometry (gated on all living cells). The cell colonies were trypsinized and incubated with the antibody 1ug/1.5 x 105cells in a permeabilization buffer. A PE conjugated goat anti-rabbit antibody was used as the secondary.

    See Abreview

  • Histone H3 immunogold detection in HeLa cells.
    Ulltra-thin sections of paraformaldehyde-fixed, Lowicryl K4M embedded cells were incubated sequentially with ab1791 antibody and an anti-rabbit antibody coupled to 10 nm gold particles. Notice absence of chromatin within Interchromatin Granules (IG), a reservoir of spliceosomal snRNPs, and high labeling of patches of condensed chromatin close to the nuclear envelope (arrows). Nu: nucleus, Cyt: cytoplasm.

    Image courtesy of Gerard Pierron, IGR-Villejuif, France.

    Sample : human
    Type : HeLa cells
    Fixative : either paraformaldehyde 4% or 1.6% glutaraldehyde in 0.1M Millonig’s buffer.
    Embedding : in Lowicryl 4KM at -20°C under UV.
    Ultrathin-sections deposited on formvar-coated carbonated EM-grids.
    Blocking step: 5% BSA for 30 seconds at RT.
    Primary antibody: ab1791 diluted 1/50 in PBS, for 1h at RT.
    Secondary antibody: BBI International anti-rabbit

  • ab1791 staining Histone H3 in adult mouse brain, dentate gyrus tissue sections by Immunohistochemistry (IHC-Fr - frozen sections).

    Tissue was fixed with paraformaldehyde, blocked with 1% BSA for 30 minutes at 25°C. Samples were incubated with primary antibody (1/500 TBS +TritonX-100 (0.05% Donkey serum 3%) for 12 hours at 4°C. Un-diluted ab175471 was used as the secondary antibody.

    See Abreview

References

This product has been referenced in:
  • Liu B  et al. Telomere shortening by transgenerational transmission of TNF-a-induced TERRA via ATF7. Nucleic Acids Res 47:283-298 (2019). Read more (PubMed: 30407559) »
  • Rassias DJ & Weathers PJ Dried leaf Artemisia annua efficacy against non-small cell lung cancer. Phytomedicine 52:247-253 (2019). Read more (PubMed: 30599905) »
See all 2721 Publications for this product

Customer reviews and Q&As

1-10 of 49 Q&A

Answer

Our antibody ab1791 will recognize total Histone H3 in your samples, so it will recognize the modified forms of Histone H3. Unfortunately I am unaware of the ratio of expression between the different variant forms in human cell lines.

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Answer

I am happy to let you know that we have non-rabbit anti-human Histone H3 antibodies available, which also qualify for our testing discount programme (either because of their potential suitability for IF or because we do not have an image yet).

These are:

ab10799: mouse monoclonal, suitable for IHC, so might work for ICC/IF

https://www.abcam.com/index.html?datasheet=10799 (or use the following: https://www.abcam.com/index.html?datasheet=10799).

ab128012: sheep polyclonal, suitable for ICC/IF, no image yet

https://www.abcam.com/index.html?datasheet=128012 (or use the following: https://www.abcam.com/index.html?datasheet=128012).

Also, I could recommend our favourite rabbit polyclonal ab1791. I assume the customer is performing a costaining with another rabbit antibody and thus wants to avoid a rabbit anti-Histone H3 antibody. However, I am pleased to let you know that with our Easy Link kits this problem can be solved: You can easily label any primary antibody with a conjugate of your choice and eliminate the need for secondary antibodies. Please find more information under https://www.abcam.com/Easylink.

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Answer

Please try the below given antibodies. These are excellent products and are guaranteed against their respective targets.

https://www.abcam.com/histone-h4-antibody-chip-grade-ab10158.html

https://www.abcam.com/histone-h3-antibody-chip-grade-ab1791.html

Read More

Answer

I've compiled some information below regarding potential primer sets for the ChIP antibodies, and where information was lacking (verified in ChIP by other researches), I included links to abReviews and references that used the antibody in ChIP.

1. ab817: GAPDH (+), GAD1 (-), AFM (-), Factor VIII (-)

2. ab1791: GAPDH, RPL30, ALDOA, MYO-D, SERPINA, AFM, SAT2, SATa (each batch is verified against this primer set, an example image of one batch test is given on the datasheet)

3. ab7030: was verified in ChIP by other researchers submitting data through our abReview system and specific references. Their positive and negative controls used are given through the link below, as well as a "click to contact this user" link that allows you to contact the researchers if you would like more specific primer information.
a) Abreview: (NFKBIA promoter (+), GAPDH (-)) https://www.abcam.com/HDAC3-antibody-ChIP-Grade-ab7030-reviews.html?intabreviewid=25057
b) Abreview: https://www.abcam.com/HDAC3-antibody-ChIP-Grade-ab7030-reviews.html?intabreviewid=18386
c) Specific references: https://www.abcam.com/HDAC3-antibody-ChIP-Grade-ab7030-references.html

4. ab111069 Untested in ChIP.

5. ab992: was verified in ChIP by other researchers (abReviews and specific references)
a) Specific references: https://www.abcam.com/Rad21-antibody-ChIP-Grade-ab992-references.html
b) Abreview: https://www.abcam.com/Rad21-antibody-ChIP-Grade-ab992-reviews.html?intabreviewid=14542
c) Abreview: (EBV 3+ (+), EBV1,2,4,5,6 (-), GAPDH (-), Cen (-)) https://www.abcam.com/Rad21-antibody-ChIP-Grade-ab992-reviews.html?intabreviewid=33942

Our ChIP primer sets:
https://www.abcam.com/index.html?pageconfig=resource&rid=10600

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Answer

Unfortunately the detection of phosphorylated Histone H3 at serine 10 is not something we have tested.

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Answer

The immunogen for ab1791 does have 100% homology with both human and mouse H3.1, H3.2, and H3.3.

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Question
Answer

ab1791 will recognize total histone H3, including any of the modified forms. The immunogen sequence has 100% homology to Human H3.3, H3.2, H3.1.

Read More

Answer

I can confirm that we do not know if ab1791 cross-reacts with CENP-A. This has not been tested experimentally.

The immunogen shows a 66% homology which means the antibody usually is not expected to react. But since there are a couple consecutive residues homologue to the immunogen, we cannot guarantee this.

We are already showing ab1791 to detect other H3 variant (WB image) on the datasheet when a the antibody is used in a high concentration.

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Answer

Vielen Dank für Ihre Anfrage.

Wir haben Ihre Anfrage an unser Labor weitergeleitet, und eine detaillierte Antwort erhalten, von der ich hoffe, dass Sie Ihnen weiterhelfen wird:

"1791 968898 formulation details below:

1x PBS, 0.02% (w/v) Sodium Azide. Note, there is no BSA in this batch

Not sure what type of conjugation they are doing but the Sodium Azide could potentially be affecting conjugation efficiency. This could just be dialysed out or use desalting. There is a current version of the protocol you could point them in direction of on the website right now (https://www.abcam.com/index.html?pageconfig=resource&rid=13869). There is also some sodium azide removal kits on our website if want to purchase these. Links are on protocol.

We also use a low pH to elute our abs and very similar what customer is using (we use 0.1M Glycine, pH 2.75). However, as the customer will probably know it is important to make sure you neutralise the ab asap or could denature your ab (we just use tris in our collection vials) and also buffer exchange into neutral buffer (such as PBS) within few hours of elution. Note, we have not had any reports of ab1791 being prone to precipitation at purification stage. However, there is a note on the internal datasheet that a batch precipitated when being transferred probably due to freeze thaw so I would avoid freeze/thaw as much as poss!"



Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

Benutzen Sie unsere Produkte? Schicken Sie uns einen Abreview. Verdienen Sie sich eine Belohnung!
https://www.abcam.com/abreviews

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1-10 of 49 Q&A

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