重组Anti-CD16+CD32抗体[EPR23501-203] (ab223200)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR23501-203] to CD16+CD32
- Suitable for: Flow Cyt, IP, ICC/IF, WB, IHC-P
- Reacts with: Mouse
Related conjugates and formulations
概述
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产品名称
Anti-CD16+CD32抗体[EPR23501-203]
参阅全部 CD16+CD32 一抗 -
描述
兔单克隆抗体[EPR23501-203] to CD16+CD32 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt, IP, ICC/IF, WB, IHC-Pmore details
不适用于: IHC-Fr -
种属反应性
与反应: Mouse -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: J774A.1 whole cell lysates; Mouse spleen and liver tissue lysates; His-tagged mouse CD16 recombinant protein; His-tagged mouse CD32 recombinant protein. IHC-P: Mouse spleen, liver and colon carcinoma tissue. ICC/IF: J774A.1 cells. Flow cyt: J774A.1 and mouse splenocyte cells. IP: J774A.1 whole cell lysate; Mouse spleen tissue lysate.
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常规说明
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.
性能
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形式
Liquid -
存放说明
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. -
存储溶液
Preservative: 0.01% Sodium azide
Constituents: 59.94% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR23501-203 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab223200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 |
---|---|---|
Flow Cyt |
1/500.
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IP |
1/30.
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ICC/IF |
1/500.
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WB |
1/1000. Detects a band of approximately 40-60 kDa.
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IHC-P |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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说明 |
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Flow Cyt
1/500. |
IP
1/30. |
ICC/IF
1/500. |
WB
1/1000. Detects a band of approximately 40-60 kDa. |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
靶标
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相关性
Function: Binds to the Fc region of immunoglobulins gamma. Low affinity receptor. By binding to IgG it initiates cellular responses against pathogens and soluble antigens. Promotes phagocytosis of opsonized antigens. Tissue specificity: Found on monocytes, neutrophils and eosinophil platelets. Similarity: Contains 2 Ig-like C2-type (immunoglobulin-like) domains. PTM: Phosphorylated by SRC-type Tyr-kinases such as LYN, BLK, FYN, HCK and SYK. -
细胞定位
CD32: Type I membrane protein. CD16: Attached to the membrane by a GPI anchor. Exists also as a soluble receptor, produced by a proteolytic cleavage. -
数据库链接
- Entrez Gene: 14130 Mouse
- Entrez Gene: 14131 Mouse
- SwissProt: P08101 Mouse
- SwissProt: P08508 Mouse
- SwissProt: Q5D5I8 Mouse
- SwissProt: Q5D5J5 Mouse
- SwissProt: Q7TMW9 Mouse
- SwissProt: Q9ES92 Mouse
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别名
- CD16a antibody
- CD16b antibody
- CD32 antibody
see all
图片
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All lanes : Anti-CD16+CD32 antibody [EPR23501-203] (ab223200) at 1/1000 dilution
Lane 1 : J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate
Lane 2 : bEnd.3 (mouse brain endothelioma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Observed band size: 40-60 kDa why is the actual band size different from the predicted?Blocking/Diluting buffer and concentration 5% NFDM/TBST
The molecular weight observed is consistent with what has been described in the literature (PMID: 19503602, 31172847). Negative control: bEnd.3 (PMID: 23911392).
Exposure time: Lanes 1-2: 5.5 seconds
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Flow cytometric analysis of mouse splenocytes cells labelling CD16+CD32 with ab223200 at 1/500 dilution (Right) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left).
Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Gated on viable cells.
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Flow cytometric analysis of bEnd.3 (mouse brain endothelioma, Left) / J774A.1 (mouse reticulum cell sarcoma monocyte macrophage, Right) cells labelling CD16+CD32 with ab223200 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled 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.
Negative control: bEnd.3 (PMID: 23911392).
Gated on viable cells.
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Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD16+CD32 with ab223200 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic and membranouse staining on mouse spleen (PMID: 22618994). The section was incubated with ab223200 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) cells labelling CD16+CD32 with ab223200 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing strong membranous and weak cytoplasmic staining in J774A.1 cells. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.
Negative control: bEnd.3 cell (PMID: 23911392).
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CD16+CD32 was immunoprecipitated from 0.35 mg Mouse spleen tissue lysate with ab223200 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab223200 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Mouse spleen tissue lysate 10 ug
Lane 2: ab223200 IP in Mouse spleen tissue lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab223200 in mouse spleen tissue lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
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All lanes : Anti-CD16+CD32 antibody [EPR23501-203] (ab223200) at 1/1000 dilution
Lane 1 : His-tagged mouse CD16 recombinant protein (aa 31-215), 5 uL
Lane 2 : His-tagged mouse CD32 recombinant protein (aa 30-210), 25 uL
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilutionBlocking and diluting buffer and concentration: 5% NFDM/TBST.
This product can recognize both CD16 and CD32.
The loading samples are E.coil extracts containing CD16 or CD32 recombinant protein respectively.
Exposure time: 70 seconds.
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CD16+CD32 was immunoprecipitated from 0.35 mg J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate with ab223200 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab223200 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate 10 ug
Lane 2: ab223200 IP in J774A.1 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab223200 in J774A.1 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds.
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All lanes : Anti-CD16+CD32 antibody [EPR23501-203] (ab223200) at 1/1000 dilution
Lane 1 : Mouse spleen tissue lysate
Lane 2 : Mouse liver tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
Observed band size: 40-60 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID: 19503602, 31172847).
Exposure time: 3 minutes.
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Immunohistochemical analysis of paraffin-embedded mouse colon carcinoma tissue labeling CD16+CD32 with ab223200 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the stroma in mouse colon carcinoma. The section was incubated with ab223200 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling CD16+CD32 with ab223200 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse liver. The section was incubated with ab223200 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.
Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.
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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SDS download
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Datasheet download
Certificate of Compliance
文献 (1)
ab223200 被引用在 1 文献中.
- Zhao P et al. Fetuin-A alleviates neuroinflammation against traumatic brain injury-induced microglial necroptosis by regulating Nrf-2/HO-1 pathway. J Neuroinflammation 19:269 (2022). PubMed: 36333786