Anti-HLA E 抗体 [MEM-E/02] (ab2216)
Key features and details
- Mouse monoclonal [MEM-E/02] to HLA E
- Suitable for: Flow Cyt, WB, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG1
製品の概要
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製品名
Anti-HLA E antibody [MEM-E/02]
HLA E 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [MEM-E/02] to HLA E -
由来種
Mouse -
特異性
This antibody reacts with the denaturated heavy chain of human HLA-E. It does not cross-react with HLA-A, -B, -C or -G. Specifity of the antibody was confirmed on HLA-G/HLA-E Workshop(Victoria 2002). -
アプリケーション
適用あり: Flow Cyt, WB, IHC-Pmore details -
種交差性
交差種: Human -
免疫原
Recombinant full length protein corresponding to Human HLA E.
Database link: P13747 -
ポジティブ・コントロール
- WB: A549, THP-1 and Jurkat cell lysates. IHC-P: Human tonsil tissue. Flow Cyt: HL60 cells.
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特記事項
This product has been changed from ascites to tissue culture supernatant. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.
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
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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: 0.097% Sodium azide
Constituent: PBS -
Concentration information loading...
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精製度
Protein A purified -
特記事項(精製)
Purified from TCS. Purity >95% by SDS-PAGE. -
ポリ/モノ
モノクローナル -
クローン名
MEM-E/02 -
ミエローマ
unknown -
アイソタイプ
IgG1 -
軽鎖の種類
unknown -
研究分野
関連製品
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
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ab2216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 |
---|---|---|
Flow Cyt |
Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
|
WB | (4) |
Use at an assay dependent concentration. Predicted molecular weight: 40 kDa.
|
IHC-P | (1) |
Use a concentration of 5 - 10 µg/ml.
|
特記事項 |
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Flow Cyt
Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 40 kDa. |
IHC-P
Use a concentration of 5 - 10 µg/ml. |
ターゲット情報
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関連性
HLA E belongs to the HLA class I heavy chain paralogues. This class I molecule is a heterodimer consisting of a heavy chain and a light chain (beta-2 microglobulin). The heavy chain is anchored in the membrane. HLA E binds a restricted subset of peptides derived from the leader peptides of other class I molecules. -
細胞内局在
Membrane; Single-pass type I membrane protein -
参照データベース
- Entrez Gene: 3133 Human
- Omim: 143010 Human
- SwissProt: P13747 Human
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別名
- HLA class I histocompatibility antigen E alpha chain antibody
- EA1.2 antibody
- EA2.1 antibody
see all
画像
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All lanes : Anti-HLA E antibody [MEM-E/02] (ab2216) at 1/500 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : HLA-E knockout A549 cell lysate
Lane 3 : THP-1 cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution
Predicted band size: 40 kDa
Observed band size: 40 kDaLanes 1-4: Merged signal (red and green). Green - ab2216 observed at 40 kDa. Red - loading control ab181602 observed at 36 kDa.
ab2216 Anti-HLA E antibody [MEM-E/02] was shown to specifically react with HLA E in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267080 (knockout cell lysate ab258452) was used. Wild-type and HLA E knockout samples were subjected to SDS-PAGE. ab2216 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated at room temperature for 2.5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-HLA E antibody [MEM-E/02] (ab2216) at 1/500 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : HLA-E knockout HEK-293T cell lysate
Lane 3 : THP-1 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 40 kDaLanes 1-3: Merged signal (red and green). Green - ab2216 observed at 40 kDa. Red - loading control ab52901 observed at kDa.
ab2216 Anti-HLA E antibody [MEM-E/02] was shown to specifically react with HLA E in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267231 (knockout cell lysate ab258454) was used. Wild-type and HLA E knockout samples were subjected to SDS-PAGE. ab2216 and Anti-beta Tubulin [EP1331Y] - Microtubule Marker (ab52901) were incubated overnight at 4°C at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-HLA E antibody [MEM-E/02] (ab2216) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : HLA-E knockout A549 cell lysate
Lane 3 : THP-1 cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) at 1/10000 dilution
Predicted band size: 40 kDaLanes 1-4: Merged signal (red and green). Green - ab2216 observed at 40 kDa. Red - loading control ab181602 observed at 36 kDa.
ab2216 Anti-HLA E antibody [MEM-E/02] was shown to specifically react with HLA E in wild-type A549 cells. Loss of signal was observed when knockout cell line ab267081 (knockout cell lysate ab258453) was used. Wild-type and HLA E knockout samples were subjected to SDS-PAGE. ab2216 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated at room temperature for 2.5 hours at 1 in 500 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) and Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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IHC image of ab2216 staining in human tonsil 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 ab2216, 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. -
Overlay histogram showing HL60 cells stained with ab2216 (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 (ab2216, 1 µg/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, 2 µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (18)
ab2216 は 18 報の論文で使用されています。
- Jin H & Kim HJ NK Cells Lose Their Cytotoxicity Function against Cancer Stem Cell-Rich Radiotherapy-Resistant Breast Cancer Cell Populations. Int J Mol Sci 22:N/A (2021). PubMed: 34502547
- Forneris N et al. Human-porcine MHC-I homology allows for antibody cross-reactivity. HLA 96:197-201 (2020). PubMed: 32452158
- Zeestraten EC et al. Combined analysis of HLA class I, HLA-E and HLA-G predicts prognosis in colon cancer patients. Br J Cancer 110:459-68 (2014). WB ; Human . PubMed: 24196788
- Engels CC et al. Immunological subtypes in breast cancer are prognostic for invasive ductal but not for invasive lobular breast carcinoma. Br J Cancer N/A:N/A (2014). Human . PubMed: 24937677
- Reimers MS et al. Prognostic value of HLA class I, HLA-E, HLA-G and Tregs in rectal cancer: a retrospective cohort study. BMC Cancer 14:486 (2014). IHC-P ; Human . PubMed: 24997850