Anti-Histone H1.5 抗体 - ChIP Grade (ab18208)
Key features and details
- Rabbit polyclonal to Histone H1.5 - ChIP Grade
- Suitable for: ChIP, WB, IHC-P, ICC/IF
- Reacts with: Human, Recombinant fragment
- Isotype: IgG
製品の概要
-
製品名
Anti-Histone H1.5 antibody - ChIP Grade
Histone H1.5 一次抗体 製品一覧 -
製品の詳細
Rabbit polyclonal to Histone H1.5 - ChIP Grade -
由来種
Rabbit -
アプリケーション
適用あり: ChIP, WB, IHC-P, ICC/IFmore details -
種交差性
交差種: Human, Recombinant fragment -
免疫原
Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Histone H1.5.
Immunogen の所有権に関して -
ポジティブ･コントロール
- Recombinant histone H1.5
-
特記事項
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. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
バッファー
pH: 7.40
Preservative: 0.02% 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... -
精製度
Immunogen affinity purified -
ポリ/モノ
ポリクローナル -
アイソタイプ
IgG -
研究分野
関連製品
-
Compatible Secondaries
-
Isotype control
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ab18208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 2 µg for 25 µg of chromatin.
|
|
| WB | (4) |
Use a concentration of 0.5 µg/ml. Detects a band of approximately 32 kDa (predicted molecular weight: 32 kDa).
|
| IHC-P |
1/800. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
|
| ICC/IF |
Use a concentration of 5 µg/ml.
|
| 特記事項 |
|---|
|
ChIP
Use 2 µg for 25 µg of chromatin. |
|
WB
Use a concentration of 0.5 µg/ml. Detects a band of approximately 32 kDa (predicted molecular weight: 32 kDa). |
|
IHC-P
1/800. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
|
ICC/IF
Use a concentration of 5 µg/ml. |
ターゲット情報
-
機能
Histones H1 are necessary for the condensation of nucleosome chains into higher order structures. -
配列類似性
Belongs to the histone H1/H5 family.
Contains 1 H15 (linker histone H1/H5 globular) domain. -
細胞内局在
Nucleus. Chromosome. - Information by UniProt
-
参照データベース
- Entrez Gene: 3009 Human
- Omim: 142711 Human
- SwissProt: P16401 Human
- Unigene: 131956 Human
-
別名
- H1 antibody
- H1 histone family member 5 antibody
- H1.5 antibody
see all
画像
-
Western blot - Anti-Histone H1.5 antibody - ChIP Grade (ab18208)This image is courtesy of Dr Albert Jordan and Monica Sancho, Center for Genomic Regulation (CRG). Recombinant H1 isoforms courtesy of Dr Nicole Happel.All lanes : Anti-Histone H1.5 antibody - ChIP Grade (ab18208) at 0.5 µg/ml
Lane 1 : Recombinant histone H1
Lane 2 : Recombinant histone H1.1
Lane 3 : Recombinant histone H1.2
Lane 4 : Recombinant histone H1.3
Lane 5 : Recombinant histone H1.4
Lane 6 : Recombinant histone H1.5
Predicted band size: 32 kDa
Observed band size: 32 kDa -
ChIP - Anti-Histone H1.5 antibody - ChIP Grade (ab18208)Chromatin was prepared from HeLa 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 ab18208 (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.
-
Immunocytochemistry/ Immunofluorescence - Anti-Histone H1.5 antibody - ChIP Grade (ab18208)ICC/IF image of ab18208 stained MCF7 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 (ab18208, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879 Alexa Fluor® 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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Histone H1.5 antibody - ChIP Grade (ab18208)IHC image of ab18208 staining in human skin formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab18208, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
Western blot - Anti-Histone H1.5 antibody - ChIP Grade (ab18208)All lanes : Anti-Histone H1.5 antibody - ChIP Grade (ab18208) at 0.5 µg/ml
Lane 1 : Histone H1 Rec
Lane 2 : Histone H1.1 Rec
Lane 3 : Histone H1.2 Rec
Lane 4 : Histone H1.3 Rec
Lane 5 : Histone H1.4 Rec
Lane 6 : Histone H1.5 Rec
Lysates/proteins at 0.1 µg per lane.
Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution
Predicted band size: 32 kDa
Observed band size: 34,37 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteThis 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 ab18208 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.
プロトコール
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (8)
ab18208 は 8 報の論文で使用されています。
- Olins AL et al. Hyperosmotic stress: in situ chromatin phase separation. Nucleus 11:1-18 (2020). PubMed: 31924112
- Teif VB et al. Linker histone epitopes are hidden by in situ higher-order chromatin structure. Epigenetics Chromatin 13:26 (2020). PubMed: 32505195
- Sang B et al. Ras-AKT signaling represses the phosphorylation of histone H1.5 at threonine 10 via GSK3 to promote the progression of glioma. Artif Cells Nanomed Biotechnol 47:2882-2890 (2019). PubMed: 31307224
- Gould TJ et al. Defining the epichromatin epitope. Nucleus 8:625-640 (2017). PubMed: 28960120
- Wang L et al. Intracellular CD24 disrupts the ARF-NPM interaction and enables mutational and viral oncogene-mediated p53 inactivation. Nat Commun 6:5909 (2015). PubMed: 25600590
- Momeni M et al. Immunohistochemical detection of promyelocytic leukemia zinc finger and histone 1.5 in uterine leiomyosarcoma and leiomyoma. Reprod Sci 21:1171-6 (2014). PubMed: 24784718
- Laserna EJ et al. Proteomic analysis of phosphorylated nuclear proteins underscores novel roles for rapid actions of retinoic acid in the regulation of mRNA splicing and translation. Mol Endocrinol 23:1799-814 (2009). PubMed: 19812389
- Sancho M et al. Depletion of human histone H1 variants uncovers specific roles in gene expression and cell growth. PLoS Genet 4:e1000227 (2008). WB ; Human . PubMed: 18927631