重组Anti-GFAP抗体[EPR1034Y] -小鼠IgG2a (Chimeric) - BSA and Azide free (ab279302)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [EPR1034Y] to GFAP - Chimeric – BSA and Azide free
- Suitable for: WB, ICC, IHC-P, Flow Cyt (Intra), IHC-Fr, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-GFAP抗体[EPR1034Y] -小鼠IgG2a (Chimeric) - BSA and Azide free
参阅全部 GFAP 一抗 -
描述
小鼠单克隆抗体[EPR1034Y] to GFAP - Chimeric – BSA and Azide free -
宿主
Mouse -
经测试应用
适用于: WB, ICC, IHC-P, Flow Cyt (Intra), IHC-Fr, IPmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- IHC-P: Human cerebral cortex tissue. WB: Human, mouse and rat brain tissue lysate. IHC-Fr: Mouse cerebrum tissue. Flow Cyt (intra): Rat primary neural/glia cells. IP: Rat brain tissue lysate. ICC: mouse primary neural/glia cell
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常规说明
ab279302 is the carrier free version of ab279290.
This mouse monoclonal chimeric antibody has been engineered from a RabMAb parent antibody (ab68428). By necessity, some rabbit sequence is retained as part of the variable domain. When multiplexing with other rabbit-derived antibodies, using cross absorbed Fc-reactive secondary antibodies are recommended.
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.
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: 100% 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
EPR1034Y -
同种型
IgG2a -
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ab279302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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WB |
Use at an assay dependent concentration.
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ICC |
1/50.
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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IHC-Fr |
Use at an assay dependent concentration.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
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IP |
Use at an assay dependent concentration.
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说明 |
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WB
Use at an assay dependent concentration. |
ICC
1/50. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
Use at an assay dependent concentration. |
IHC-Fr
Use at an assay dependent concentration. Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20). |
IP
Use at an assay dependent concentration. |
靶标
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功能
GFAP, a class-III intermediate filament, is a cell-specific marker that, during the development of the central nervous system, distinguishes astrocytes from other glial cells. -
组织特异性
Expressed in cells lacking fibronectin. -
疾病相关
Defects in GFAP are a cause of Alexander disease (ALEXD) [MIM:203450]. Alexander disease is a rare disorder of the central nervous system. It is a progressive leukoencephalopathy whose hallmark is the widespread accumulation of Rosenthal fibers which are cytoplasmic inclusions in astrocytes. The most common form affects infants and young children, and is characterized by progressive failure of central myelination, usually leading to death usually within the first decade. Infants with Alexander disease develop a leukoencephalopathy with macrocephaly, seizures, and psychomotor retardation. Patients with juvenile or adult forms typically experience ataxia, bulbar signs and spasticity, and a more slowly progressive course. -
序列相似性
Belongs to the intermediate filament family. -
翻译后修饰
Phosphorylated by PKN1. -
细胞定位
Cytoplasm. Associated with intermediate filaments. - Information by UniProt
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数据库链接
- Entrez Gene: 2670 Human
- Entrez Gene: 14580 Mouse
- Entrez Gene: 24387 Rat
- Omim: 137780 Human
- SwissProt: P14136 Human
- SwissProt: P03995 Mouse
- SwissProt: P47819 Rat
- Unigene: 514227 Human
see all -
别名
- wu:fb34h11 antibody
- ALXDRD antibody
- cb345 antibody
see all
图片
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This data was developed using ab279290, the same antibody clone in a different buffer formulation.
Immunocytochemical analysis of 4% PFA-fixed, 0.1% Triton X-100 permeabilized mouse neural/glia tissue labeling GFAP with ab279290 at 1/50 (16 µg/ml) dilution followed by ab98736 Goat Anti-Mouse IgG (DyLight® 488) pre-adsorbed at 1/1000 dilution (Green). Positive staining on mouse primary astrocytes is observed. The nuclear counterstain was DAPI (Blue).
Counterstain: ab207165 anti-GFAP rabbit mAb at 1/1000 dilution with secondary antibody ab150088 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) pre-adsorbed at 1/1000 dilution.
Negative controls: ab279290 with secondary ab150080 at 1/500 dilution; and ab207165 at 1/1000 dilution with secondary ab98376 at 1/1000 dilution
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All lanes : Anti-GFAP antibody [EPR1034Y] - Mouse IgG2a (Chimeric) (ab279290) at 1/1000 dilution
Lane 1 : Human brain tissue lysate
Lane 2 : Mouse brain tissue lysate
Lane 3 : Rat brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/5000 dilution
Observed band size: 40-50 kDa why is the actual band size different from the predicted?This data was produced using ab279290, the same clone in a different formulation.
Blocking/Dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with the literature (PMID: 824020, 2294, 6340792).
Exposure times: Lane 1: 3.25 seconds; Lane 2: 48 seconds; Lane 3: 10 seconds.
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This data was produced using ab279290, the same clone in a different formulation.
IHC image of GFAP staining in a section of formalin-fixed paraffin-embedded normal human cerebral cortex performed on a Leica BONDTM system using the standard protocol F.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab279290, 1µg/ml, for 15 mins at room temperature. A rabbit anti-mouse IgG2a, was added for 8 mins at room temperature and detected using an HRP conjugated goat anti-rabbit compact polymer system. DAB was used as the chromogen.
The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
This data was produced using ab279290, the same clone in a different formulation.
Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cerebrum tissue labeling GFAP with ab279290 at 1/500 (1.634 µg/ml) dilution followed by ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse cerebrum is observed. The nuclear counterstain was DAPI (Blue).
Secondary antibody control: Secondary antibody is ab150113 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488)at 1/1000 dilution.
Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
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This data was produced using ab279290, the same clone in a different formulation.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized rat primary neural/glia cells labelling GFAP with ab279290 at 1/1000 dilution (0.1µg)/ Right compared with a Mouse monoclonal IgG isotype control/ Left. Goat Anti-Mouse IgG (Alexa Fluor® 647, ab150119) at 1/2000 dilution was used as the secondary antibody.
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This data was produced using ab279290, the same clone in a different formulation.
GFAP was immunoprecipitated from 0.35 mg rat brain tissue lysate 10 µg with ab279290 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab279290 at 1/1000 dilution. mouse IgG for IP (HRP) (ab131368) was used at 1/5000 dilution.
Lane 1: Rat brain tissue lysate 10µg.
Lane 2: ab279290 IP in rat brain tissue lysate.
Lane 3: Mouse monoclonal IgG2a (ab18413) instead of ab279290 in rat brain tissue lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 23 seconds.
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
文献 (1)
ab279302 被引用在 1 文献中.
- Jiang Y et al. Gypenoside-14 Reduces Depression via Downregulation of the Nuclear Factor Kappa B (NF-kB) Signaling Pathway on the Lipopolysaccharide (LPS)-Induced Depression Model. Pharmaceuticals (Basel) 16:N/A (2023). PubMed: 37631068