Anti-PRAME 抗体 [EPR20330] (ab219650)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20330] to PRAME
- Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-PRAME antibody [EPR20330]
PRAME 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR20330] to PRAME -
由来種
Rabbit -
特異性
PRAME is expressed in malignant cells, including leukaemias, Hodgkin's lymphoma, breast cancer, and primary and metastatic melanomas.
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アプリケーション
適用あり: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details -
種交差性
交差種: Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: MeWo and A-375 whole cell lysates; Human ovary cancer and testis lysates. IHC-P: Human melanoma, breast carcinoma and human testis tissue. ICC/IF: MeWo and A-375 cells. Flow Cyt (intra): MeWo cells, K562 cells. IP: MeWo whole cell lysate.
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特記事項
PRAME (PReferentially expressed Antigen in MElanoma) is a tumor-associated antigen and is a member of the family of cancer testis antigens (CTA). PRAME is expressed in malignant cells, including leukaemias, Hodgkin's lymphoma, breast cancer, and primary and metastatic melanomas. For more information, please refer to PMID: 27441500. PRAME has low or no expression in normal tissues except for in testis, ovary, placenta, adrenals and endometrium. For more information, please refer to PMID: 9047241.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
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 long term. Avoid freeze / thaw cycle. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR20330 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Compatible Secondaries
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Immunohistochemistry kits
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Isotype control
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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ab219650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 |
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WB |
1/1000. Detects a band of approximately 57 kDa (predicted molecular weight: 57 kDa).
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IHC-P |
1/100 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Recommend ab219650 incubation at +4°C overnight. |
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ICC/IF |
1/500.
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IP |
1/30.
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Flow Cyt (Intra) |
1/500.
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特記事項 |
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WB
1/1000. Detects a band of approximately 57 kDa (predicted molecular weight: 57 kDa). |
IHC-P
1/100 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Recommend ab219650 incubation at +4°C overnight. |
ICC/IF
1/500. |
IP
1/30. |
Flow Cyt (Intra)
1/500. |
ターゲット情報
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機能
Functions as a transcriptional repressor, inhibiting the signaling of retinoic acid through the retinoic acid receptors RARA, RARB and RARG. Prevents retinoic acid-induced cell proliferation arrest, differentiation and apoptosis. -
組織特異性
Expressed in testis. Detected in samples of kidney, brain and skin. -
配列類似性
Belongs to the PRAME family.
Contains 4 LRR (leucine-rich) repeats. -
細胞内局在
Nucleus. Cell membrane. - Information by UniProt
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参照データベース
- Entrez Gene: 23532 Human
- Omim: 606021 Human
- SwissProt: P78395 Human
- Unigene: 30743 Human
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別名
- 4930534P07Rik antibody
- Cancer/testis antigen 130 antibody
- CT130 antibody
see all
画像
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Flow cytometry overlay histogram showing left K562 positive cells and right negative HEK293 stained with ab219650 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilised with 0.1 % PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab219650) (1x 106 in 100μl at 0.008μg/ml (1/267500)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
This antibody gave a positive signal in K562 Fixed with 80% methanol (5 min) / permeabilised with 0.1 % PBS-Triton X-100 for 15 min under the same conditions.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)
Immunohistochemical analysis of paraffin-embedded human melanoma tissue labeling PRAME with ab219650 at 1/500 dilution, followed by LeicaDS9800 (Bond™ Polymer Refine Detection). Sections were counter stained with Hematoxylin. Antigen retrieval was heat mediated with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Nuclear staining on human melanoma. The section was incubated with ab219650 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.Secondary antibody only control: Used PBS instead of primary antibody.
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PRAME immunofluorescence with PRAME antibody ab219650 in K562 cells, with negative expression in Ramos cells. The cells were fixed with 4% formaldehyde (10 min), permeabilised with 0.1% 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 ab219650 at 1 µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin at 0.5 µg/ml. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150119, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 647), pre-adsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.
This product also work with 100% methanol (5 min) fixation under the same testing conditions.
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PRAME flow cytometry with PRAME antibody ab219650 of 4% paraformaldehyde-fixed and 90% Methanol-permeabilised MeWo (Human malignant melanoma cell line) cells labeling PRAME with ab219650 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-PRAME antibody [EPR20330] (ab219650) at 1/1000 dilution
Lane 1 : Human ovary cancer lysate
Lane 2 : A-375 (Human malignant melanoma cell line) whole cell lysate
Lane 3 : Human testis lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 57 kDa
Observed band size: 57 kDaPRAME western blot with PRAME antibody ab219650.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1: 3 minutes; Lane 2: 5 seconds; Lane 3: 1 minute.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized MeWo (Human malignant melanoma cell line) cells labeling PRAME with ab219650 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing mostly nuclear staining on MeWo cells.
The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)
Immunohistochemical analysis of paraffin-embedded human testis tissue labeling PRAME with ab219650 at 1/500 dilution, followed by LeicaDS9800 (Bond™ Polymer Refine Detection). Sections were counter stained with Hematoxylin. Antigen retrieval was heat mediated with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Nuclear staining on human testis. The section was incubated with ab219650 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.Secondary antibody only control: Used PBS instead of primary antibody.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling PRAME with ab219650 at 1/500 dilution, followed by LeicaDS9800 (Bond™ Polymer Refine Detection). Sections were counter stained with Hematoxylin. Antigen retrieval was heat mediated with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Nuclear staining on human breast carcinoma. The section was incubated with ab219650 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.Secondary antibody only control: Used PBS instead of primary antibody.
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PRAME was immunoprecipitated from 0.35 mg of MeWo (Human malignant melanoma cell line) whole cell lysate with ab219650 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab219650 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: MeWo whole cell lysate 10 μg (Input).
Lane 2: ab219650 IP in MeWo whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab219650 in MeWo whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
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PRAME western blot with PRAME antibody ab219650.
Different batches of ab219650 were tested on MeWo (Human malignant melanoma fibroblast) whole cell lysate at 0.1 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 50 kDa.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A-375 (Human malignant melanoma cell line) cells labeling PRAME with ab219650 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing mostly nuclear staining on A-375 cells.
The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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Anti-PRAME antibody [EPR20330] (ab219650) at 1/1000 dilution + MeWo (Human malignant melanoma cell line) whole cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/4000 dilution
Predicted band size: 57 kDa
Observed band size: 57 kDa
Exposure time: 15 secondsPRAME western blot with PRAME antibody ab219650.
Blocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)
Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling PRAME with ab219650 at 1/500 dilution, followed by LeicaDS9800 (Bond™ Polymer Refine Detection). Sections were counter stained with Hematoxylin. Antigen retrieval was heat mediated with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Negative control: No staining on human stomach. The section was incubated with ab219650 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)
Immunohistochemical analysis of paraffin-embedded human breast tissue labeling PRAME with ab219650 at 1/500 dilution, followed by LeicaDS9800 (Bond™ Polymer Refine Detection). Sections were counter stained with Hematoxylin. Antigen retrieval was heat mediated with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Negative control: No staining on human breast. The section was incubated with ab219650 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling PRAME with ab219650 at 1/500 dilution, followed by LeicaDS9800 (Bond™ Polymer Refine Detection). Sections were counter stained with Hematoxylin. Antigen retrieval was heat mediated with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Negative control: No staining on human tonsil. The section was incubated with ab219650 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PRAME antibody [EPR20330] (ab219650)
Tissue microarray (TMA) analysis using PRAME EPR20330 (ab219650).
This table provides a detailed overview of positive PRAME staining (tick mark) and negative PRAME staining (cross mark) per sample type tested. PRAME is expressed in metastatic melanoma (PMID: 30045064). PRAME has low or no expression in normal tissues except in testis, ovary, placenta, adrenals and endometrium (PMID: 30045064).
The sections were pre-treated using Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab219650 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
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
Certificate of Compliance
参考文献 (29)
ab219650 は 29 報の論文で使用されています。
- Chen YP et al. PRAME is a useful marker for the differential diagnosis of melanocytic tumours and histological mimics. Histopathology 82:285-295 (2023). PubMed: 36200756
- Jung JM et al. Performance of PRAME immunohistochemistry compared with that of c-Kit, c-Myc, or cyclin D1 for the diagnosis of acral melanocytic tumors. Pathol Int 73:27-36 (2023). PubMed: 36468840
- Schossig P et al. Target Selection for T-Cell Therapy in Epithelial Ovarian Cancer: Systematic Prioritization of Self-Antigens. Int J Mol Sci 24:N/A (2023). PubMed: 36768616
- Yu L et al. HDAC5-mediated PRAME regulates the proliferation, migration, invasion, and EMT of laryngeal squamous cell carcinoma via the PI3K/AKT/mTOR signaling pathway. Open Med (Wars) 18:20230665 (2023). PubMed: 36910848
- Hedrich V et al. PRAME Is a Novel Target of Tumor-Intrinsic Gas6/Axl Activation and Promotes Cancer Cell Invasion in Hepatocellular Carcinoma. Cancers (Basel) 15:N/A (2023). PubMed: 37173882