Anti-Histone H3 (tri methyl K27) 抗体 [mAbcam 6002] - ChIP Grade (ab6002)

製品の概要

  • 製品名
    Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade
    Histone H3 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [mAbcam 6002] to Histone H3 (tri methyl K27) - ChIP Grade
  • 特異性
    This antibody is specific for histone H3 tri-methylated at K27. The antibody is blocked in Western blot by tri methyl K27 peptide and slightly by di methyl K27 peptide (there is <12% cross reactivity with di methyl K27 as determined by ELISA). 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 the peptide blocking assay blot below).
  • アプリケーション
    適用あり: ChIP/Chip, ChIP, IHC-P, Flow Cyt, ELISA, WB, ICC, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Cow, Human, Arabidopsis thaliana, Drosophila melanogaster, Plants, Zebrafish, Rhesus monkey, Rice
    交差が予測される動物種: Rabbit, Chicken, Xenopus laevis, Chinese hamster
  • 免疫原

    Synthetic peptide within Human Histone H3 aa 1-100 (tri methyl K27) conjugated to keyhole limpet haemocyanin (Sulfosuccinimidyl 4-N-maleimidomethyl-cyclohexane-1-carboxylate (Sulfo-SMCC)). The exact sequence is proprietary. Clones were positively screened by ELISA against the immunising peptide. Clones were negatively screened against both the unmodified peptide and also against tri methyl K9 Histone H3 peptide.
    (Peptide available as ab1782)

  • 特記事項

    Western blot protocol advice:

    For the blocking step, we recommend using 3% milk for 1 hour at room temperature. This step may need to be optimized depending on your experimental conditions.

    This antibody clone [mAbcam 6002] is manufactured by Abcam.
    We have the following conjugates available:

    Anti-Histone H3 (tri methyl K27) antibody (Alexa Fluor® 488) [mAbcam 6002] (ab205728)
    Anti-Histone H3 (tri methyl K27) antibody (Alexa Fluor® 647) [mAbcam 6002] (ab205729)

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
ChIP/Chip Use at an assay dependent concentration.
ChIP Use 5-10 µg for 25 µg of chromatin.
IHC-P Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab18392 - Mouse monoclonal IgG3, is suitable for use as an isotype control with this antibody.

ELISA Use a concentration of 0.025 - 1 µg/ml.
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 Human Histone H3 (tri methyl K27) peptide (ab1782).

Blocking: we recommend using 3% milk block for 1 hour at room temperature. This step may need to be optimized for your experiments.

ICC Use at an assay dependent concentration.
ICC/IF Use a concentration of 5 µg/ml.

ターゲット情報

  • 機能
    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.
  • 配列類似性
    Belongs to the histone H3 family.
  • 発生段階
    Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
  • 翻訳後修飾
    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.
  • 細胞内局在
    Nucleus. Chromosome.
  • Information by UniProt
  • 参照データベース
    see all
  • 別名
    • HIST1 cluster, H3J antibody
    • Histone gene cluster 1, H3 histone family, member E antibody
    • Histone gene cluster 1, H3G antibody
    • H3 histone family, member J antibody
    • HIST1 cluster, H3E antibody
    • HIST1 cluster, H3I antibody
    • Histone gene cluster 1, H3C 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, member T antibody
    • H3 histone, family 3A antibody
    • H3.1 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
    • H3/t antibody
    • H31_HUMAN antibody
    • H3F1K antibody
    • H3F3 antibody
    • H3F3A antibody
    • H3FA antibody
    • H3FB antibody
    • H3FC antibody
    • H3FD antibody
    • H3FF antibody
    • H3FH antibody
    • H3FI antibody
    • H3FJ antibody
    • H3FK antibody
    • H3FL antibody
    • HIST1 cluster, H3A antibody
    • HIST1 cluster, H3B antibody
    • HIST1 cluster, H3C antibody
    • HIST1 cluster, H3D antibody
    • HIST1 cluster, H3F antibody
    • HIST1 cluster, H3G antibody
    • HIST1 cluster, H3H 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 3, H3 antibody
    • histone cluster 1 H3 family member a antibody
    • histone cluster 1 H3 family member b antibody
    • histone cluster 1 H3 family member c antibody
    • histone cluster 1 H3 family member d antibody
    • histone cluster 1 H3 family member e antibody
    • histone cluster 1 H3 family member f antibody
    • histone cluster 1 H3 family member g antibody
    • histone cluster 1 H3 family member h antibody
    • histone cluster 1 H3 family member i antibody
    • histone cluster 1 H3 family member j 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 gene cluster 1, H3 histone family, member A antibody
    • Histone gene cluster 1, H3 histone family, member B antibody
    • Histone gene cluster 1, H3 histone family, member C antibody
    • Histone gene cluster 1, H3 histone family, member D antibody
    • Histone gene cluster 1, H3 histone family, member F antibody
    • Histone gene cluster 1, H3 histone family, member G antibody
    • Histone gene cluster 1, H3 histone family, member H antibody
    • Histone gene cluster 1, H3 histone family, member I antibody
    • Histone gene cluster 1, H3 histone family, member J antibody
    • Histone gene cluster 1, H3A antibody
    • Histone gene cluster 1, H3B antibody
    • Histone gene cluster 1, H3D antibody
    • Histone gene cluster 1, H3E antibody
    • Histone gene cluster 1, H3F antibody
    • Histone gene cluster 1, H3H antibody
    • Histone gene cluster 1, H3I antibody
    • Histone gene cluster 1, H3J antibody
    • Histone H 3 antibody
    • Histone H3.1 antibody
    • histone H3.1t 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

画像

  • In mice, the X-inactivation process is reversed naturally by X-reactivation in blastocysts and germ cells and in culture in pluripotent stem cells. The image shows late blastocyst-stage mouse embryos consisting of three cell types: epiblast (NANOG-positive, cyan), primitive endoderm (GATA4-positive, red) and trophectoderm (CDX2-positive, green). The inactive X-chromosome (H3K27me3-positive, yellow dots) is reactivated only in the epiblast (cells without yellow spots), which will form the embryo. The germ cell factor PRDM14 and the long noncoding RNA Tsix collaborate during the X-reactivation process in blastocysts and pluripotent stem cells and thereby link epigenetic with cellular reprogramming events.

    Image is courtesy of Bernhard Payer, runner-up of the immunofluorescence imaging competition 2017.

  • 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.
        

  • 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

  • All batches of ab6002 are tested in ELISA against peptides to different Histone H3 modifications. Results show strong binding to Histone H3 - tri methyl K27 immunising peptide (ab1782), indicating that this antibody specifically recognises the Histone H3 - tri methyl K27 modification. Weak binding is also detected against the Histone H3 - di methyl K27 modification (<12%) (ab1781).

    ab17163 - Histone H3 - unmodified

    ab1340 - Histone H3 - mono methyl K4

    ab7768 - Histone H3 - di methyl K4

    ab1342 - Histone H3 - tri methyl K4

    ab1771 - Histone H3 - mono methyl K9

    ab1772 - Histone H3 - di methyl K9

    ab1773 - Histone H3 - tri methyl K9

    ab1780 - Histone H3 - mono methyl K27

    ab1781 - Histone H3 - di methyl K27

    ab1782 - Histone H3 - tri methyl K27

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

    Lysates/proteins at 0.25 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 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 : 3 minutes

    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 3% milk before being incubated with ab6002 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • Lanes 1 - 3 : Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002) at 1 µg/ml (2% BSA)
    Lanes 4 - 6 : Anti-Histone H3 (tri methyl K27) antibody [mAbcam 6002] - ChIP Grade (ab6002) at 1 µg/ml (3% MILK)

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 2 : EED-/- mouse ES Whole Cell Lysate
    Lane 3 : WT mouse ES Whole Cell Lysate
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
    Lane 5 : EED-/- mouse ES Whole Cell Lysate
    Lane 6 : WT mouse ES Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 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 : 12 minutes

    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 (lanes 1-3) and 3% milk (lanes 4-6) before being incubated with ab6002 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • 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.

  • 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.

  • 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.
  • Paraformaldehyde-fixed, paraffin-embedded human invasive breast carcinoma tissue stained for Histone H3 (tri methyl K27) using ab6002 at 1/200 dilution in immunohistochemical analysis.

    See Abreview

参考文献

This product has been referenced in:
  • Oittinen M  et al. Polycomb Repressive Complex 2 Enacts Wnt Signaling in Intestinal Homeostasis and Contributes to the Instigation of Stemness in Diseases Entailing Epithelial Hyperplasia or Neoplasia. Stem Cells 35:445-457 (2017). WB, ChIP . Read more (PubMed: 27570105) »
  • Ye Y  et al. Chromatin remodeling during in vivo neural stem cells differentiating to neurons in early Drosophila embryos. Cell Death Differ 24:409-420 (2017). ChIP ; Drosophila melanogaster . Read more (PubMed: 27858939) »

See all 381 Publications for this product

レビューと Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Human Cell lysate - nuclear (K562 CML cell line)
Negative control
Rabbit IgG
Specification
K562 CML cell line
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: Formaldehyde
Positive control
GATA1 antibody
Username

Abcam user community

Verified customer

投稿 Sep 29 2017

Application
ChIP
Sample
Human Cell lysate - whole cell (esophagus)
Negative control
IgG
Specification
esophagus
Detection step
Real-time PCR
Type
Native ChIP (N-ChIP)
Positive control
polymerase ii
Username

Abcam user community

Verified customer

投稿 Aug 14 2017

Application
ChIP
Sample
Human Cell lysate - whole cell (Human Epithelial OVCAR cell line)
Negative control
IgG
Specification
Human Epithelial OVCAR cell line
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: Formaldehyde (1%)
Positive control
-
Username

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投稿 Jul 05 2017

Application
ChIP
Sample
Human Cell lysate - nuclear (Adipose stromal cells)
Negative control
no ab / Input
Specification
Adipose stromal cells
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: PFA, 1%
Positive control
H3K27ac
Username

Abcam user community

Verified customer

投稿 Jun 02 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (adipose stem cells)
Permeabilization
Yes - Triton 0.25%
Specification
adipose stem cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
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投稿 Mar 07 2017

Application
Immunohistochemistry (Resin sections)
Sample
Astatotilapia burtoni Tissue sections (Brain preoptic area LRWhite embedded)
Specification
Brain preoptic area LRWhite embedded
Username

Sebastian Alvarado

Verified customer

投稿 May 03 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Vascular smooth muscle cell)
Permeabilization
Yes - NP40
Specification
Vascular smooth muscle cell
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 24°C
Fixative
Formaldehyde
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Verified customer

投稿 Feb 17 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (Human medulloblastoma cells)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Specification
Human medulloblastoma cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Feb 01 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Astatotilapia burtoni Tissue sections (Brain thalamus)
Antigen retrieval step
None
Permeabilization
Yes - 0.2% TritonX
Specification
Brain thalamus
Blocking step
10% normal serum with 1% BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
Username

Sebastian Alvarado

Verified customer

投稿 Dec 18 2015

Application
Western blot
Sample
Mouse Tissue lysate - whole (Whole brain tissue lysate)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (4-12% Bis-Tris Gel, MES Running Buffer)
Loading amount
15 µg
Specification
Whole brain tissue lysate
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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投稿 Jun 15 2015

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

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