重组Anti-CD14抗体[EPR21847] (ab221678)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21847] to CD14
- Suitable for: IHC-Fr, IP, Flow Cyt, ICC/IF, WB
- Reacts with: Mouse
Related conjugates and formulations
概述
-
产品名称
Anti-CD14抗体[EPR21847]
参阅全部 CD14 一抗 -
描述
兔单克隆抗体[EPR21847] to CD14 -
宿主
Rabbit -
经测试应用
适用于: IHC-Fr, IP, Flow Cyt, ICC/IF, WBmore details -
种属反应性
与反应: Mouse -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
阳性对照
- WB: J774A.1 and RAW264.7 whole cell lysate; mouse lymph node and placenta lysates. ICC/IF: J774A.1 and RAW 264.7 cells. Flow cyt: RAW 264.7 cells, C57 BL/6 mouse bone marrow cells. IP: RAW 264.7 whole cell lysate; IHC-Fr: Mouse spleen tissue.
-
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
-
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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
-
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR21847 -
同种型
IgG -
研究领域
相关产品
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab221678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 |
---|---|---|
IHC-Fr |
Use at an assay dependent concentration.
|
|
IP |
1/30.
|
|
Flow Cyt |
1/500.
|
|
ICC/IF |
1/1000.
|
|
WB |
1/1000. Detects a band of approximately 50-55 kDa (predicted molecular weight: 39 kDa).
|
说明 |
---|
IHC-Fr
Use at an assay dependent concentration. |
IP
1/30. |
Flow Cyt
1/500. |
ICC/IF
1/1000. |
WB
1/1000. Detects a band of approximately 50-55 kDa (predicted molecular weight: 39 kDa). |
靶标
-
功能
Cooperates with MD-2 and TLR4 to mediate the innate immune response to bacterial lipopolysaccharide (LPS). Acts via MyD88, TIRAP and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the inflammatory response. Up-regulates cell surface molecules, including adhesion molecules. -
组织特异性
Expressed strongly on the surface of monocytes and weakly on the surface of granulocytes; also expressed by most tissue macrophages. -
序列相似性
Contains 11 LRR (leucine-rich) repeats. -
翻译后修饰
N- and O- glycosylated. O-glycosylated with a core 1 or possibly core 8 glycan. -
细胞定位
Cell membrane. - Information by UniProt
-
数据库链接
- Entrez Gene: 12475 Mouse
- SwissProt: P10810 Mouse
-
别名
- CD 14 antibody
- CD_antigen=CD14 antibody
- CD14 antibody
see all
图片
-
Flow cytometry staining of C57 BL/6 mouse bone marrow cells with ab221678 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were incubated for 30 min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab221678 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 µl at 10.0 μg/ml (1/215)) for 30min on ice. The cells were simultaneously stained with Ly6G.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice
Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.
-
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse spleen tissue labeling CD14 with ab221678 at 1/50 (10.62 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) at 1/1000 dilution (Green). Positive staining on mouse spleen. is observed. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor ®; 488) at 1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
-
Flow cytometric analysis of RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cell line labeling CD14 with ab221678 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells incubated with secondary antibody only) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)(ab150077) at 1/2000 dilution was used as the secondary antibody.
Gated on viable cells.
-
CD14 was immunoprecipitated from 0.35 mg RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate with ab221678 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221678 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 μg (Input).
Lane 2: ab221678 IP in RAW 264.7 whole cell lysate(+).
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab221678 in RAW 264.7 whole cell lysate (-).Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 secondsThe molecular mass observed is consistent with the literature (PMID: 9502426; PMID:7513013)
-
All lanes : Anti-CD14 antibody [EPR21847] (ab221678) at 1/1000 dilution
Lane 1 : J774A.1 (mouse reticulum cell sarcoma monocyte macrophage), whole cell lysate at 10 µg
Lane 2 : Mouse lymph node lysate at 20 µg
Lane 3 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 10 µg
Lane 4 : Mouse placenta lysate at 10 µg
Secondary
Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Developed using the ECL technique.
Predicted band size: 39 kDa
Observed band size: 50-55 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: Lane 1: 6 seconds; Lane 2: 3 minutes; Lane 3: 10 seconds; Lane 4: 81 seconds.
The molecular mass observed is consistent with the literature (PMID: 9502426; PMID: 7513013).
-
Immunofluorescent analysis of 100% methanol-fixed RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labeling CD14 with ab221678 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)(ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining on RAW 264.7 cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (re
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488)(ab150077) secondary at 1/1000 dilution.
-
Immunofluorescent analysis of 100% methanol-fixed J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) cells labeling CD14 with ab221678 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining on J774A.1 cell line. The nuclear counter stain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary at 1/1000 dilution.
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
-
SDS download
-
Datasheet download
Certificate of Compliance
文献 (8)
ab221678 被引用在 8 文献中.
- Zhao L et al. Dendritic cell-mediated chronic low-grade inflammation is regulated by the RAGE-TLR4-PKCβ1 signaling pathway in diabetic atherosclerosis. Mol Med 28:4 (2022). PubMed: 35062863
- Pei S et al. Lysophosphatidic Acid Receptor 3 Suppress Neutrophil Extracellular Traps Production and Thrombosis During Sepsis. Front Immunol 13:844781 (2022). PubMed: 35464399
- Lin C et al. PTEN-induced kinase 1 enhances the reparative effects of bone marrow mesenchymal stromal cells on mice with renal ischaemia/reperfusion-induced acute kidney injury. Hum Cell 35:1650-1670 (2022). PubMed: 35962179
- Yang R et al. Telocytes-derived extracellular vesicles alleviate aortic valve calcification by carrying miR-30b. ESC Heart Fail 8:3935-3946 (2021). PubMed: 34165260
- Li Y et al. Polarization of rheumatoid macrophages is regulated by the CDKN2B-AS1/ MIR497/TXNIP axis. Immunol Lett 239:23-31 (2021). PubMed: 34418490
- Woo J et al. High-throughput and high-efficiency sample preparation for single-cell proteomics using a nested nanowell chip. Nat Commun 12:6246 (2021). PubMed: 34716329
- Rao L et al. Hybrid cellular membrane nanovesicles amplify macrophage immune responses against cancer recurrence and metastasis. Nat Commun 11:4909 (2020). PubMed: 32999291
- Sun Q et al. Bactericidal/Permeability-Increasing Protein Improves Cognitive Impairment in Diabetic Mice via Blockade of the LPS-LBP-TLR4 Signaling Pathway. Front Physiol 11:718 (2020). PubMed: 33643054