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リコンビナント

Anti-GFP 抗体 [EPR14104] - Chicken IgY (Chimeric) (ab300643)

Submit a review Submit a question References (1)
Western blot - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
  • Flow Cytometry (Intracellular) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
  • Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Chicken monoclonal [EPR14104] to GFP - Chimeric
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P
  • Reacts with: Species independent

製品の概要

  • 製品名

    Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric)
    GFP 一次抗体 製品一覧

  • 製品の詳細

    Chicken monoclonal [EPR14104] to GFP - Chimeric

  • 由来種

    Chicken chimera

  • アプリケーション

    適用あり: Flow Cyt (Intra), ICC/IF, WB, IHC-Pmore details

  • 種交差性

    交差種: Species independent

  • 免疫原

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: GFP transfected 293 whole cell lysate. IHC-P: HEK-293T transfected with GFP expression vector containing a His tag. ICC/IF: 293/GFP cells. HEK-293T cells transfected with Firefly Luciferase expression vector containing a GFP tag. Flow Cyt (Intra): 293T transfected with GFP tagged Firefly Luciferase over expression vector.

  • 特記事項

    This chicken monoclonal chimeric antibody has been engineered from a RabMAb parent antibody (ab183734). 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.

    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.

製品の特性

  • 製品の状態

    Liquid

  • 保存方法

    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.

  • バッファー

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • 精製度

    Thiophilic Chromatography

  • ポリ/モノ

    モノクローナル

  • クローン名

    EPR14104

  • アイソタイプ

    IgY

  • 研究分野

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab300643の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
Flow Cyt (Intra)
1/500.
ICC/IF
1/50.
WB
1/1000. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).
IHC-P
1/10000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
特記事項
Flow Cyt (Intra)
1/500.
ICC/IF
1/50.
WB
1/1000. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).
IHC-P
1/10000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ターゲット情報

  • 関連性

    Function: Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca2+ -activated photoprotein aequorin.

    Subunit structure: Monomer.

    Tissue specificity: Photocytes.

    Post-translational modification: Contains a chromophore consisting of modified amino acid residues. The chromophore is formed by autocatalytic backbone condensation between Ser-65 and Gly-67, and oxidation of Tyr-66 to didehydrotyrosine. Maturation of the chromophore requires nothing other than molecular oxygen.

    Biotechnological use: Green fluorescent protein has been engineered to produce a vast number of variously colored mutants, fusion proteins, and biosensors. Fluorescent proteins and its mutated allelic forms, blue, cyan and yellow have become a useful and ubiquitous tool for making chimeric proteins, where they function as a fluorescent protein tag. Typically they tolerate N- and C-terminal fusion to a broad variety of proteins. They have been expressed in most known cell types and are used as a noninvasive fluorescent marker in living cells and organisms. They enable a wide range of applications where they have functioned as a cell lineage tracer, reporter of gene expression, or as a measure of protein-protein interactions. Can also be used as a molecular thermometer, allowing accurate temperature measurements in fluids. The measurement process relies on the detection of the blinking of GFP using fluorescence correlation spectroscopy.

    Sequence similarities: Belongs to the GFP family.

    Biophysicochemical properties: Absorption: Abs(max)=395 nm
    Exhibits a smaller absorbance peak at 470 nm. The fluorescence emission spectrum peaks at 509 nm with a shoulder at 540 nm.

  • 別名

    • GFP antibody
    • Green fluorescent protein antibody

画像

  • Western blot - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
    Western blot - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
    All lanes : Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643) at 1/1000 dilution

    Lane 1 : Non-transfected 293T (human embryonic kidney epithelial cell) whole cell lysate
    Lane 2 : GFP transfected 293 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Chicken IgY H&L (HRP) (ab6877) at 1/20000 dilution

    Predicted band size: 27 kDa
    Observed band size: 27 kDa



    Blocking / Diluting buffer and concentration: 5% NFDM/TBST

    Exposure time: 3.25 seconds

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)

    Immunohistochemical analysis of paraffin-embedded (A) HEK-293T transfected with GFP expression vector containing a his tag. (B) Wild type HEK-293T cells labeling GFP with ab300643 at 1/10000 dilution (0.996 µg/ml) followed by ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. Positive staining on HEK-293T transfected with GFP expression vector containing a his tag (Image: A); No staining on Wild type HEK-293T cells (Image: B). The section was incubated with ab300643 for 30 mins at room temperature followed by anti-chicken IgG antibody (ab97136) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)

    Immunohistochemical analysis of paraffin-embedded human tonsil labeling GFP with ab300643 at 1/10000 dilution (0.996 µg/ml) followed by ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. No staining on human tonsil. The section was incubated with ab300643 for 30 mins at room temperature followed by anti-chicken IgG antibody (ab97136) for 8 mins during the Leica DS9800 kit staining procedure. 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 LeicaDS9800 (BOND™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)

    Immunohistochemical analysis of paraffin-embedded mouse spleen labeling GFP with ab300643 at 1/10000 dilution (0.996 µg/ml) followed by ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. No staining on mouse spleen. The section was incubated with ab300643 for 30 mins at room temperature followed by anti-chicken IgG antibody (ab97136) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)

    Immunohistochemical analysis of paraffin-embedded rat spleen labeling GFP with ab300643 at 1/10000 dilution (0.996 µg/ml) followed by ready to use Leica DS9800 (BOND™ Polymer Refine Detection) kit. No staining on rat spleen. The section was incubated with ab300643 for 30 mins at room temperature followed by anti-chicken IgG antibody (ab97136) for 8 mins during the LeicaDS9800 kit staining procedure. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
    Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)

    Immunofluorescent analysis of 4% Paraformaldehyde fixed, 0.1% Triton X-100 permeabilized 293/GFP (human embryonic kidney epithelial cell) cells labeling GFP with ab300643 at 1/50 dilution followed by ab150175 Goat Anti-Chicken IgY H&L (Alexa Fluor® 647) preadsorbed secondary antibody at 1/1000 (2 μg/ml) dilution (red). Confocal image showing positive staining in 293/GFP cells. The nuclear counter stain is DAPI (blue).

  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
    Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)

    Immunofluorescent analysis of 4% Paraformaldehyde fixed, 0.1% Triton X-100 permeabilized HEK293T (human embryonic kidney epithelial cells) labeling GFP with ab300643 at 1/50 dilution followed by ab150175 Goat Anti-Chicken IgY H&L (Alexa Fluor® 647) preadsorbed secondary antibody at 1/1000 (2 μg/ml) dilution (red). Confocal image showing positive staining in HEK-293T cells transfected with a Firefly Luciferase expression vector containing a GFP tag. The nuclear counter stain is DAPI (blue).

  • Flow Cytometry (Intracellular) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
    Flow Cytometry (Intracellular) - Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)

    Flow Cytometry (Intracellular) analysis of 293T transfected with GFP tagged Firefly Luciferase over expression vector labeling GFP (right) with ab300643 at a 1/500 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat anti-Chicken IgY H&L (Alexa Fluor® 647, ab150175) was used as the secondary antibody at a 1/5000 dilution.  Chicken IgY Isotype control. (left)

  • Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)
    Anti-GFP antibody [EPR14104] - Chicken IgY (Chimeric) (ab300643)

プロトコール

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

参考文献 (1)

ab300643 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab300643 は 1 報の論文で使用されています。

  • Diniz F  et al. Acetyl-CoA is a key molecule for nephron progenitor cell pool maintenance. Nat Commun 14:7733 (2023). PubMed: 38007516

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