重组Anti-CD276抗体[EPNCIR122] - BSA and Azide free (ab256585)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPNCIR122] to CD276 - BSA and Azide free
- Suitable for: IHC-Fr, WB, IP, Flow Cyt, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
概述
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产品名称
Anti-CD276抗体[EPNCIR122] - BSA and Azide free
参阅全部 CD276 一抗 -
描述
兔单克隆抗体[EPNCIR122] to CD276 - BSA and Azide free -
宿主
Rabbit -
经测试应用
适用于: IHC-Fr, WB, IP, Flow Cyt, ICC/IFmore details
不适用于: IHC-P -
种属反应性
与反应: Mouse, Human -
免疫原
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: LnCaP, CHO-K1 cells lysates transfected with human and mouse CD276. Flow Cyt: HEK293 and THP1 cells. ICC/IF: HEK293 cells.
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常规说明
ab256585 is the carrier-free version of ab134161.
This antibody was developed as part of a collaboration between Epitomics, the National Cancer Institute's Center for Cancer Research and the lab of Brad St. Croix.
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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
存储溶液
pH: 7.2
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPNCIR122 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Anti-CD276 antibody [EPNCIR122] (ab134161)
- Anti-CD276 antibody [EPNCIR122] - Low endotoxin, Azide free (ab209895)
- Alexa Fluor® 488 Anti-CD276 antibody [EPNCIR122] (ab218280)
- Alexa Fluor® 594 Anti-CD276 antibody [EPNCIR122] (ab218282)
- Alexa Fluor® 555 Anti-CD276 antibody [EPNCIR122] (ab218283)
- Alexa Fluor® 647 Anti-CD276 antibody [EPNCIR122] (ab300754)
- PE Anti-CD276 antibody [EPNCIR122] (ab314419)
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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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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab256585于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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IHC-Fr |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Detects a band of approximately 110 kDa (predicted molecular weight: 57 kDa).
Please check the parent abID, ab134161, for more information on dilutions. |
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IP |
Use at an assay dependent concentration.
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Flow Cyt |
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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ICC/IF |
Use a concentration of 1 µg/ml.
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说明 |
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IHC-Fr
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Detects a band of approximately 110 kDa (predicted molecular weight: 57 kDa). Please check the parent abID, ab134161, for more information on dilutions. |
IP
Use at an assay dependent concentration. |
Flow Cyt
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
Use a concentration of 1 µg/ml. |
靶标
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功能
May participate in the regulation of T-cell-mediated immune response. May play a protective role in tumor cells by inhibiting natural-killer mediated cell lysis as well as a role of marker for detection of neuroblastoma cells. May be involved in the development of acute and chronic transplant rejection and in the regulation of lymphocytic activity at mucosal surfaces. Could also play a key role in providing the placenta and fetus with a suitable immunological environment throughout pregnancy. Both isoform 1 and isoform 2 appear to be redundant in their ability to modulate CD4 T-cell responses. Isoform 2 is shown to enhance the induction of cytotoxic T-cells and selectively stimulates interferon gamma production in the presence of T-cell receptor signaling. -
组织特异性
Ubiquitous but not detectable in peripheral blood lymphocytes or granulocytes. Weakly expressed in resting monocytes. Expressed in dendritic cells derived from monocytes. Expressed in epithelial cells of sinonasal tissue. Expressed in extravillous trophoblast cells and Hofbauer cells of the first trimester placenta and term placenta. -
序列相似性
Belongs to the immunoglobulin superfamily. BTN/MOG family.
Contains 2 Ig-like C2-type (immunoglobulin-like) domains.
Contains 2 Ig-like V-type (immunoglobulin-like) domains. -
细胞定位
Membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 80381 Human
- Entrez Gene: 102657 Mouse
- Omim: 605715 Human
- SwissProt: Q5ZPR3 Human
- SwissProt: Q8VE98 Mouse
- Unigene: 77873 Human
- Unigene: 5356 Mouse
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别名
- 4Ig B7 H3 antibody
- 4Ig-B7-H3 antibody
- AU016588 antibody
see all
图片
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All lanes : Anti-CD276 antibody [EPNCIR122] (ab134161) at 1/1000 dilution
Lane 1 : LLC (Mouse Lewis lung carcinoma cell) whole cell lysate (boiled)
Lane 2 : LLC (Mouse Lewis lung carcinoma cell) whole cell lysate (unboiled)
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 57 kDa
Observed band size: 45-66 kDa why is the actual band size different from the predicted?
Exposure time: 180 secondsBlocking and dilution buffer: 5% NFDM/TBST.
We recommend not to boil the samples after lysis to get desired WB results.
Mouse CD276 (B7-H3) has a molecular weight about 45–66 kDa, depending on the glycosylation levels [PMID: 34366684].This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134161).
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Overlay histogram showing THP-1 (human monocytic leukemia monocyte) cells stained with ab134161 (red line). The cells were fixed with 4% paraformaldehyde and then permeabilized with 90% methanol. The cells were incubated with the antibody (ab134161) at 1/80 dilution. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution. Isotype control antibody (black line) was rabbit monoclonal IgG (ab172730) used under the same conditions. Unlabelled sample (blue line) was also used as a control.
Performed using purified antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134161).
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This data was developed using the same antibody clone in a different buffer formulation (ab134161).
ab134161 staining CD276 in wild-type HEK293 cells (top panel) and CD276 knockout HEK293 cells (ab266658) (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized 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 with ab134161 at 1μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8). -
Overlay histogram showing THP1 cells stained with ab134161 (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% human serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab134161, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134161).
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CD276 was immunoprecipitated from 10 ug of HEK 293 (human embryonic kidney epithelial cell) whole cell lysate with ab134161 at 1/40 diltuion. Western blot was permformed from the immunoprecipitate using ab 134161 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/500 dilution.
Lane 1: HEK 293 whole cell lysate 10 ug (Input).
Lane 2: ab134161 IP in HEK 293 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab134161 in HEK 293 whole cell lysate
.Blocking/dilution buffer: 5% NFDM/TBSTPerformed using purified antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134161).
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Immunohistochemistry of mouse MC38 colon liver metastasis. Staining CD276 with ab134161 (green). Normal tissue (N)/tumor tissue (T) margins are indicated by a white dash.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab134161).
实验方案
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.
数据表及文件
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Datasheet download
Certificate of Compliance
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