Anti-GFP抗体[6AT316] (ab38689)

概述

  • 产品名称
    Anti-GFP抗体[6AT316]
    参阅全部 GFP 一抗
  • 描述
    小鼠单克隆抗体[6AT316] to GFP
  • 特异性
    Detects both GFP and YFP by Western Blotting.
  • 经测试应用
    适用于: IHC-Fr, ELISA, WB, IHC-Pmore details
  • 免疫原

    Full length native protein (purified) corresponding to GFP.
    Database link: P42212

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.
  • 存储溶液
    Preservative: 0.09% Sodium Azide
    Constituents: PBS
  • Concentration information loading...
  • 纯度
    Protein G purified
  • 纯化说明
    This antibody was purified by protein G and Ni-NTA nickel affinity chromatography, followed by dialysis against PBS.
  • 克隆
    单克隆
  • 克隆编号
    6AT316
  • 同种型
    IgG1
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab38689 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
IHC-Fr 1/500.
ELISA 1/1000.
WB 1/4000. Predicted molecular weight: 27 kDa.
IHC-P 1/50 - 1/100.

靶标

  • 相关性
    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.
  • 别名
    • GFP antibody
    • Green fluorescent protein antibody

图片

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse skin tissue labelling GFP with ab38689. Tissue was fixed with formaldehyde and blocked with 3% BSA for 0.5 hour at 38°C; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/25) for 1 hours at 37°C. A peroxidase-conjugated goat anti-mouse polyclonal (ready to use) was used as the secondary antibody. -ve shows negative staining of the antibody in a wild type mouse, whereas +ve shows positive staining in a mouse expressing GFP under the LGR5 promoter.

  • Immunohistochemistry (Frozen sections) analysis of E10.5 mouse embryo tissue sections labeling GFP with ab38689 at 1/200 dilution. The tissue was fixed with paraformaldehyde and permeabilized with PBS / 0.5% v/v Triton X-100. The tissue was incubated with ab38689 in 1% BSA + 5% goat serum + PBST for 12 hours at 25°C. A polyclonal goat anti-mouse Alexa Fluor® 568 secondary antibody was used at 1/1000 dilution.

    See Abreview

  • Anti-GFP antibody [6AT316] (ab38689) at 1/4000 dilution + GFP recombinant protein lysate at 35 µg

    Secondary
    Peroxidase-conjugated goat anti-mouse IgG (H+L) at 1/5000 dilution

    Predicted band size : 27 kDa

    Incubation time was overnight at 4°C. Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-GFP antibody [6AT316] (ab38689)

    Lane 1 : GFP at 1 µg
    Lane 2 : GFP at 0.1 µg
    Lane 3 : GFP at 0.01 µg
    Lane 4 : YFP at 1 µg
    Lane 5 : YFP at 0.1 µg
    Lane 6 : YFP at 0.01 µg


    Predicted band size : 27 kDa
    Observed band size : 27 kDa
    GFP and YFP were expressed in bacteria.
  • Formalin-fixed and paraffin-embedded human cancer tissue reacted with ab38689, which was peroxidase-conjugated to the secondary antibody, followed by AEC staining.
  • ab38689 staining GFP in Chicken spinal cord tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 20°C. Samples were incubated with primary antibody (1/500 in PBS) for 16 hours at 4°C. A Cy2®-conjugated Donkey anti-mouse monoclonal (1/100) was used as the secondary antibody.

    See Abreview

文献

This product has been referenced in:
  • Chuah JA  et al. Gene introduction into the mitochondria of Arabidopsis thaliana via peptide-based carriers. Sci Rep 5:7751 (2015). WB . Read more (PubMed: 25583214) »
  • Derghal A  et al. Leptin is required for hypothalamic regulation of miRNAs targeting POMC 3'UTR. Front Cell Neurosci 9:172 (2015). Read more (PubMed: 25999818) »

See all 14 Publications for this product

客户评价及客户问答

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (pancreas)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mm sodium citrate PH 6
Permeabilization
No
Specification
pancreas
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 18°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Jul 13 2017

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (HeLa Cells)
Total protein in input
1e+007 cells
Treatment
Transfected with EGFP 48 hrs
Immuno-precipitation step
Protein G
Specification
HeLa Cells
Username

Abcam user community

Verified customer

提交于 Oct 20 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
NOVOCASTRA protein block as blocking agent as blocking agent as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Sample
Mouse Tissue sections (E10.5 mouse embryo)
Specification
E10.5 mouse embryo
Permeabilization
Yes - PBS / 0.5% v/v Triton X-100 (Sigma)
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Mar 20 2014

The epitope of the antibody has not been mapped, and it has not been tested with eGFP, so we cannot guarantee it detects eGFP. However, EGFP differs from GFP by only a few amino acids, there is a great chance the antibody will detect eGFP.

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Chicken Tissue sections (Spinal cord)
Specification
Spinal cord
Fixative
Paraformaldehyde
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Dr. A Klar

Verified customer

提交于 Apr 12 2011

PROCEDURE: Solution Preparation PBS, pH7.4: 10 mM Na2HPO4, 1.8 mM KH2PO4, 50 mM NaCl, 2.7 mM KCl Blocking buffers: 5% BSA/0.5% Tween-20 in 1X PBS. Protocol 1. Deparaffinize and rehydrate sections as follows: 3 x 3 min with xylene; 3 x 2...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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