Anti-mH2A1 抗体 [14G7] (ab91528)
Key features and details
- Mouse monoclonal [14G7] to mH2A1
- Suitable for: ChIP, WB, Flow Cyt (Intra), ICC/IF
- Knockout validated
- Reacts with: Human
- Isotype: IgG2
製品の概要
-
製品名
Anti-mH2A1 antibody [14G7]
mH2A1 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [14G7] to mH2A1 -
由来種
Mouse -
アプリケーション
適用あり: ChIP, WB, Flow Cyt (Intra), ICC/IFmore details -
種交差性
交差種: Human -
免疫原
Recombinant full length protein corresponding to Human mH2A1.
-
ポジティブ・コントロール
- This antibody gave a positive signal in the following whole cell lysates: HeLa; HepG2; MCF7
-
特記事項
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
製品の特性
-
製品の状態
Liquid -
保存方法
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles. -
バッファー
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading...
-
精製度
Protein G purified -
ポリ/モノ
モノクローナル -
クローン名
14G7 -
アイソタイプ
IgG2 -
軽鎖の種類
lambda -
研究分野
関連製品
-
Compatible Secondaries
-
Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab91528の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
---|---|---|
ChIP |
Use at an assay dependent concentration.
|
|
WB |
Use a concentration of 10 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa).
|
|
Flow Cyt (Intra) |
Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
|
ICC/IF | (1) |
Use a concentration of 1 µg/ml.
|
特記事項 |
---|
ChIP
Use at an assay dependent concentration. |
WB
Use a concentration of 10 µg/ml. Detects a band of approximately 40 kDa (predicted molecular weight: 40 kDa). |
Flow Cyt (Intra)
Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use a concentration of 1 µg/ml. |
ターゲット情報
-
機能
Variant histone H2A which replaces conventional H2A in a subset of nucleosomes where it represses transcription. 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. Involved in stable X chromosome inactivation. Inhibits the binding of transcription factors and interferes with the activity of remodeling SWI/SNF complexes. Inhibits histone acetylation by EP300 and recruits class I HDACs, which induces an hypoacetylated state of chromatin. In addition, isoform 1, but not isoform 2, binds ADP-ribose and O-acetyl-ADP-ribose, and may be involved in ADP-ribose-mediated chromatin modulation. -
組織特異性
Ubiquitous. -
配列類似性
Contains 1 histone H2A domain.
Contains 1 Macro domain. -
翻訳後修飾
Monoubiquitinated at either Lys-116 or Lys-117. May also be polyubiquitinated. Ubiquitination is mediated by the CUL3/SPOP E3 complex and does not promote proteasomal degradation. Instead, it is required for enrichment in inactive X chromosome chromatin. -
細胞内局在
Nucleus. Chromosome. Enriched in inactive X chromosome chromatin and in senescence-associated heterochromatin. - Information by UniProt
-
参照データベース
- Entrez Gene: 9555 Human
- Omim: 610054 Human
- SwissProt: O75367 Human
- Unigene: 420272 Human
- Unigene: 599225 Human
-
別名
- Core histone macro h2a.1 antibody
- Core histone macro-H2A.1 antibody
- H2A histone family member Y antibody
see all
画像
-
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: mH2A1 knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)
Lane 4: HepG2 whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab91528 observed at 40 kDa. Red - loading control, ab176560, observed at 50 kDa.
ab91528 was shown to specifically recognize mH2A1 in wild-type HAP1 cells. No band was observed when mH2A1 knockout cells were examined. Wild-type and mH2A1 knockout samples were subjected to SDS-PAGE. ab91528 and ab176560 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4°C at 10 µg/ml and 1/10,000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.
-
ab91528 staining mH2A1 in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab91528 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 4% paraformaldehyde (10 min).Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
-
Overlay histogram showing HeLa cells stained with ab91528 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab91528, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
-
Chromatin was prepared from SK-OV-3 cells according to the Abcam Dual-X-ChIP protocol*. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab91528 (red), or 5 µg of mouse normal IgG1 ab18443 (gray) and 25 µl of Protein A/G Dyna beads. 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 commercial primers from Paper PMID: 22589551
*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
All lanes : Anti-mH2A1 antibody [14G7] (ab91528) at 10 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 40 kDa
Observed band size: 40 kDa
Additional bands at: 14 kDa, 18 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 2 minutes
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (0)
ab91528 は論文での使用が確認できていません。