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重组RabMAb

重组Anti-14-3-3 zeta抗体[EPR27044-19] - BSA and Azide free (ab317439)

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Western blot - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
  • Western blot - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
  • Western blot - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
  • Immunocytochemistry/ Immunofluorescence - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
  • Immunocytochemistry/ Immunofluorescence - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
  • Flow Cytometry (Intracellular) - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
  • Immunoprecipitation - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
  • Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR27044-19] to 14-3-3 zeta - BSA and Azide free
  • Suitable for: ICC/IF, WB, Flow Cyt (Intra), IP
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Unconjugated

概述

  • 产品名称

    Anti-14-3-3 zeta抗体[EPR27044-19] - BSA and Azide free
    参阅全部 14-3-3 zeta 一抗

  • 描述

    兔单克隆抗体[EPR27044-19] to 14-3-3 zeta - BSA and Azide free

  • 宿主

    Rabbit

  • 经测试应用

    适用于: ICC/IF, WB, Flow Cyt (Intra), IPmore details
    不适用于: IHC-P

  • 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.

  • 阳性对照

    • WB: 293T, 293T transfected with siRNA specifically targeting 14-3-3 zeta whole 5, HeLa, MCF7, A549, SH-SY5Y, Neuro-2a, RAW 264.7, C6, Mouse brain, Rat brain, Human lung cancer, Human cerebellum and His-tagged full-length human 14-3-3 zeta / YWHAZ recombinant protein. ICC/IF: 293T and RAW 264.7 cells. Flow Cyt (Intra): 293T cell. IP: 293T whole cell lysate.

  • 常规说明

    ab317439 is the carrier-free version of ab317438.

    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
    For more information see here.

    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

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab317439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
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
说明
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration.
Flow Cyt (Intra)
Use at an assay dependent concentration.
IP
Use at an assay dependent concentration.
应用说明
Is unsuitable for IHC-P.

靶标

  • 功能

    Adapter protein implicated in the regulation of a large spectrum of both general and specialized signaling pathways. Binds to a large number of partners, usually by recognition of a phosphoserine or phosphothreonine motif. Binding generally results in the modulation of the activity of the binding partner.

  • 序列相似性

    Belongs to the 14-3-3 family.

  • 翻译后修饰

    The delta, brain-specific form differs from the zeta form in being phosphorylated (By similarity). Phosphorylation on Ser-184 by MAPK8; promotes dissociation of BAX and translocation of BAX to mitochondria. Phosphorylation on Ser-58 by PKA; disrupts homodimerization and heterodimerization with YHAE and TP53. This phosphorylation appears to be activated by sphingosine. Phosphorylation on Thr-232; inhibits binding of RAF1.

  • 细胞定位

    Cytoplasm. Melanosome. Located to stage I to stage IV melanosomes.
  • Information by UniProt

  • 数据库链接

    see all

  • 别名

    • 14 3 3 delta antibody
    • 14 3 3 protein zeta/delta antibody
    • 14 3 3 protein/cytosolic phospholipase A2 antibody
    • 14 3 3 zeta antibody
    • 14-3-3 protein zeta/delta antibody
    • 1433Z_HUMAN antibody
    • Epididymis luminal protein 4 antibody
    • Epididymis secretory protein Li 3 antibody
    • HEL S 3 antibody
    • HEL4 antibody
    • KCIP-1 antibody
    • KCIP1 antibody
    • MGC111427 antibody
    • MGC126532 antibody
    • MGC138156 antibody
    • Phospholipase A2 antibody
    • Protein kinase C inhibitor protein 1 antibody
    • Tyrosine 3 monooxygenase/tryptophan 5 monooxygenase activation protein, delta polypeptide antibody
    • Tyrosine 3 monooxygenase/tryptophan 5 monooxygenase activation protein, zeta antibody
    • Tyrosine 3 monooxygenase/tryptophan 5 monooxygenase activation protein, zeta polypeptide antibody
    • Tyrosine 3/tryptophan 5 monooxygenase activation protein, zeta polypeptide antibody
    • YWHAD antibody
    • YWHAZ antibody
    see all

图片

  • Western blot - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
    Western blot - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
    All lanes : Anti-14-3-3 zeta antibody [EPR27044-19] (ab317438) at 1/1000 dilution

    Lane 1 : 293T (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 5 µg
    Lane 2 : 293T transfected with siRNA specifically targeting 14-3-3 zeta whole cell lysate at 5 µg
    Lane 3 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg
    Lane 4 : MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg
    Lane 5 : A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg
    Lane 6 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg
    Lane 7 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg
    Lane 8 : RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg
    Lane 9 : C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Observed band size: 27 kDa why is the actual band size different from the predicted?


    Exposure time: 8 seconds


    This data was developed using ab317438, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control  (ab181602) staining at 1/200000 dilution.

  • Western blot - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
    Western blot - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
    All lanes : Anti-14-3-3 zeta antibody [EPR27044-19] (ab317438) at 1/1000 dilution

    Lane 1 : Mouse brain tissue lysate
    Lane 2 : Rat brain tissue lysate
    Lane 3 : Human lung cancer tissue lysate
    Lane 4 : Human cerebellum tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lanes 1-2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lanes 3-4 : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

    Observed band size: 27 kDa why is the actual band size different from the predicted?



    This data was developed using ab317438, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    Lanes 1-2 are applied with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 and lanes 3-4 are applied with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

    In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control  (ab181602) staining at 1/200000 dilution.

    Exposure time: Lanes 1-2: 8 seconds; lanes 3-4: 1 second

  • Western blot - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
    Western blot - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
    All lanes : Anti-14-3-3 zeta antibody [EPR27044-19] (ab317438) at 1/1000 dilution

    Lane 1 : His-tagged full-length human 14-3-3 zeta / YWHAZ recombinant protein 10 ng
    Lane 2 : His-tagged full-length human 14-3-3 epsilon / YWHAE recombinant protein 10 ng
    Lane 3 : His-tagged full-length human 14-3-3 sigma / YWHAS recombinant protein 10 ng
    Lane 4 : His-tagged full-length human 14-3-3 eta / YWHAH recombinant protein 10 ng
    Lane 5 : His-tagged full-length human 14-3-3 gamma / YWHAG recombinant protein 10 ng
    Lane 6 : His-tagged full-length human 14-3-3 tau / YWHAQ recombinant protein 10 ng
    Lane 7 : His-tagged full-length human 14-3-3 beta / YWHAB recombinant protein 10 ng

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Observed band size: 27 kDa why is the actual band size different from the predicted?


    Exposure time: 81 seconds


    This data was developed using ab317438, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: 5% NFDM/TBST.

    This antibody does not cross-react with human 14-3-3 epsilon, 14-3-3 sigma, 14-3-3 eta, 14-3-3 gamma, 14-3-3 tau and 14-3-3 beta.

    In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade  (ab213204) staining at 1/5000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
    Immunocytochemistry/ Immunofluorescence - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)

    This data was developed using ab317438, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized 293T (human embryonic kidney epithelial cell) cells labelling 14-3-3 zeta with ab317438 at 1/50 (9.86 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

    Confocal image showing cytoplasmic staining in 293T cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
    Immunocytochemistry/ Immunofluorescence - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)

    This data was developed using ab317438, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labelling 14-3-3 zeta with ab317438 at 1/50 (9.86 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2 ug/ml) dilution (Green).

    Confocal image showing cytoplasmic staining in RAW 264.7 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2 ug/ml) dilution.

  • Flow Cytometry (Intracellular) - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
    Flow Cytometry (Intracellular) - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)

    This data was developed using ab317438, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized 293T (human embryonic kidney epithelial cell) cells labelling 14-3-3 zeta with ab317438 at 1/500 dilution (0.1ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

    Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
    Immunoprecipitation - Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)

    This data was developed using ab317438, the same antibody clone in a different buffer formulation.

    14-3-3 zeta was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) whole cell lysate with ab317438 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317438 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: 293T (human embryonic kidney epithelial cell) whole cell lysate
    Lane 2: ab317438 IP in 293T (human embryonic kidney epithelial cell) whole cell lysate
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab317438 in 293T whole cell lysate

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 6 seconds.

  • Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)
    Anti-14-3-3 zeta antibody [EPR27044-19] - BSA and Azide free (ab317439)

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.

Click here to view the general protocols

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