Anti-CD27 抗体 [EPR8569] - BSA and Azide free (ab256583)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR8569] to CD27 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-CD27 antibody [EPR8569] - BSA and Azide free
CD27 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR8569] to CD27 - BSA and Azide free -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), WB, ICC/IF, IHC-Pmore details -
種交差性
交差種: Human -
免疫原
Synthetic peptide. 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: Raji, Ramos and NAMALWA cell lysates and human lymph node and fetal spleen tissue lysates. IHC-P: Human stomach and tonsil tissues. ICC/IF: Jurkat cells. Flow Cyt (intra): Ramos cells, Human PBMCs.
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特記事項
ab256583 is the carrier-free version of ab131254.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
バッファー
pH: 7.2
Constituent: PBS -
キャリア・フリー
はい -
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
EPR8569 -
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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Conjugation kits
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Isotype control
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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ab256583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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
Use at an assay dependent concentration. Detects a band of approximately 55 kDa (predicted molecular weight: 29 kDa).
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ICC/IF |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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特記事項 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Detects a band of approximately 55 kDa (predicted molecular weight: 29 kDa). |
ICC/IF
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ターゲット情報
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機能
Receptor for CD70/CD27L. May play a role in survival of activated T-cells. May play a role in apoptosis through association with SIVA1. -
組織特異性
Found in most T-lymphocytes. -
配列類似性
Contains 3 TNFR-Cys repeats. -
翻訳後修飾
Phosphorylated and O-glycosylated. -
細胞内局在
Membrane. - Information by UniProt
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参照データベース
- Entrez Gene: 939 Human
- Omim: 186711 Human
- SwissProt: P26842 Human
- Unigene: 355307 Human
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別名
- CD 27 antibody
- CD27 antibody
- CD27 antigen antibody
see all
画像
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).
Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab131254 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were fixed and permeabilised with BD Cytofix/Cytoperm™ for 20 min. PBMCs were incubated for 30 min at 22°C in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab131254 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 µl at 0.04 μg/ml (1/52750)) for 30 min at 4°C . The cells were simultaneously stained with CD4.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 4°C
Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.
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Flow Cytometry analysis of Ramos cells labelling CD27 with purified ab131254 at 1/300 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).
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All lanes : Anti-CD27 antibody [EPR8569] (ab131254) at 1/1000 dilution
Lane 1 : Wild-type Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 2 : CD27 knockout Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 3 : Ramos (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 4 : Human brain tissue lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 29 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab131254).
Lanes 1 - 4: Merged signal (red and green). Green - ab131254 observed at 35 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab131254 was shown to react with CD27 in wild-type Raji cells in western blot with loss of signal observed in CD27 knockout sample. Wild-type and CD27 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab131254 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle showing negative staining with purified ab131254 at 1/1800. Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ab214880, a Goat Anti-Rabbit IgG H&L (HRP polymer) was used as the secondary antibody (1/500).
Counterstained with hematoxylin.
Negative control: No staining on human skeletal muscle. The section was incubated with ab131254 at 4°C overnight.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD27 with purified ab131254 at 1/1500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).
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This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).
ab131254 staining CD27 in Raji cells, with negative expression in A375 cells. The cells were fixed with 100% methanol (5 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 ab131254 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 4% formaldehyde (10 min) fixation under the same testing conditions.
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Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling CD27 with purified ab131254 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.
Control 1: primary antibody (1/500) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).
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Immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections) analysis of human stomach tissue labelling CD27 with ab131254 at a dilution of 1/100.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).
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Immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections) analysis of human tonsil tissue labelling CD27 with ab131254 at a dilution of 1/100.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).
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Equilibrium disassociation constant (KD)
Learn more about KD
Click here to learn more about KDThis data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab131254).
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
データシートおよび資料
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Datasheet download
Certificate of Compliance
参考文献 (2)
ab256583 は 2 報の論文で使用されています。
- Setayesh SM et al. Targeted single-cell proteomic analysis identifies new liquid biopsy biomarkers associated with multiple myeloma. NPJ Precis Oncol 7:95 (2023). PubMed: 37723227
- Zhou Y et al. Puerarin improves graft bone defect through microRNA-155-3p-mediated p53/TNF-a/STAT1 signaling pathway. Int J Mol Med 46:239-251 (2020). PubMed: 32377717