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RecombinantRabMAb

Recombinant Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)

Reviews (7) Submit a question References (4)
ChIP - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
  • Peptide Array - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
  • Western blot - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
  • Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR16994(2)] to Histone H3 (di methyl K36) - ChIP Grade
  • Suitable for: PepArr, ICC/IF, ChIP, IHC-P, WB
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade
    See all Histone H3 primary antibodies

  • Description

    Rabbit monoclonal [EPR16994(2)] to Histone H3 (di methyl K36) - ChIP Grade

  • Host species

    Rabbit

  • Tested applications

    Suitable for: PepArr, ICC/IF, ChIP, IHC-P, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HeLa and NIH/3T3 whole cell lysates. IHC-P: Human stomach, mouse testis and rat spleen tissues. ICC/IF: HeLa and NIH/3T3 cells. ChIP: Chromatin from HeLa cells.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab176921 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
PepArr
Use at an assay dependent concentration.
ICC/IF
1/500.
ChIP
Use 2 µg for 25 µg of chromatin.
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB (4)
1/5000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).
Notes
PepArr
Use at an assay dependent concentration.
ICC/IF
1/500.
ChIP
Use 2 µg for 25 µg of chromatin.
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/5000. Detects a band of approximately 15 kDa (predicted molecular weight: 15 kDa).

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

    see all

  • 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

  • ChIP - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
    ChIP - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)

    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 25µg of chromatin, 2µg of ab176921 (red), and 20µl of Anti rabbit IgG sepharose beads. 2μg of rabbit normal IgG was added to the beads control (grey). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Histone H3 (di methyl K36) with ab176921 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab176921 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Peptide Array - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
    Peptide Array - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)

    ab176921 was tested in Peptide array against 501 different modified and unmodified histone peptides; each peptide is printed on the array at six concentrations (each in triplicate).
     
    Circle area represents affinity between the antibody and a peptide: all antigen-containing peptides are displayed as red circles, all other peptides as blue circles. The affinity is calculated as area under curve when antibody binding values are plotted against the corresponding peptide concentration. Each circle area is normalized to the peptide with the strongest affinity.
     
    The complete dataset, including full list of all peptides and information on the position of each peptide in the diagram, can be downloaded here.

  • Western blot - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
    Western blot - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
    All lanes : Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921) at 1/5000 dilution

    Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/1000 dilution

    Predicted band size: 15 kDa
    Observed band size: 15 kDa


    Exposure time: 4 seconds


    Blocking/Dilution buffer: 5% BSA/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)

    Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling Histone H3 (di methyl K36) with ab176921 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on human stomach is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling Histone H3 (di methyl K36) with ab176921 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on mouse testis is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)

    Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling Histone H3 (di methyl K36) with ab176921 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on rat spleen is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
    Immunocytochemistry/ Immunofluorescence - Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Histone H3 (di methyl K36) with ab176921 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue).

    Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab176921 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
    -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)
    Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade (ab176921)

References (4)

Publishing research using ab176921? Please let us know so that we can cite the reference in this datasheet.

ab176921 has been referenced in 4 publications.

  • Xue W  et al. Long non-coding RNAs MACC1-AS1 and FOXD2-AS1 mediate NSD2-induced cisplatin resistance in esophageal squamous cell carcinoma. Mol Ther Nucleic Acids 23:592-602 (2021). PubMed: 33552680
  • Bressan RB  et al. Regional identity of human neural stem cells determines oncogenic responses to histone H3.3 mutants. Cell Stem Cell 28:877-893.e9 (2021). PubMed: 33631116
  • Zhou R  et al. Characterization of H3 methylation in regulating oocyte development in cyprinid fish. Sci China Life Sci N/A:N/A (2018). PubMed: 30443860
  • Streubel G  et al. The H3K36me2 Methyltransferase Nsd1 Demarcates PRC2-Mediated H3K27me2 and H3K27me3 Domains in Embryonic Stem Cells. Mol Cell 70:371-379.e5 (2018). PubMed: 29606589

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

ChIP abreview for Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade

 Inconclusive
Abreviews
abreview image
Application
ChIP
Sample
Human Cell lysate - nuclear (K562)
Specification
K562
Detection step
Other
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MR. Andrew Perez

Verified customer

Submitted Mar 24 2022

Western blot abreview for Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade

 Poor
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (T cells or lung cells)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
90 µg
Specification
T cells or lung cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 19 2022

Western blot abreview for Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade

 Inconclusive
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - nuclear (Cell line)
Gel Running Conditions
Reduced Denaturing
Loading amount
33 µg
Specification
Cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 12 2021

Western blot abreview for Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade

 Good
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - nuclear (cell line)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
57 µg
Specification
cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 29 2021

ChIP on chip abreview for Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade

 Good
Abreviews
abreview image
Application
ChIP on chip
Sample
Mouse Cell lysate - whole cell (cell line)
Specification
cell line
Detection step
Microarray
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 at room temper
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 29 2021

Western blot abreview for Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade

 Good
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - nuclear (Cell line)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
57 µg
Specification
Cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 21 2021

ChIP on chip abreview for Anti-Histone H3 (di methyl K36) antibody [EPR16994(2)] - ChIP Grade

 Good
Abreviews
abreview image
Application
ChIP on chip
Sample
Mouse Cell lysate - whole cell (Cell line)
Specification
Cell line
Detection step
Microarray
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Oct 04 2021

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