概述

  • 产品名称
  • 描述
    兔多克隆抗体to GFP
  • 宿主
    Rabbit
  • 特异性
    This antibody is reactive against all variants of Aequorea victoria GFP such as S65T-GFP, RS-GFP, YFP, CFP, RFP and EGFP.
  • 经测试应用
    适用于: IHC-FoFr, Electron Microscopy, IHC-P, IHC-Fr, IP, WB, ICC/IF, Flow Cytmore details
  • 免疫原

    Recombinant full length protein corresponding to GFP.
    Database link: P42212

  • 阳性对照
    • Pure GFP protein, or cells known to overexpress GFP.
  • 常规说明

    Please note that a mistake was made in reference 4 (Mesaeli et.al., J. Cell. Biol. 1999 Mar 8;144(5):857-68). The antibody used for immunohistochemistry on paraformaldehyde fixed tissues was the crude serum version of this antibody (Abcam ab290) and not Clontech's monoclonal as stated. This product is supplied in 25% glycerol. During freezeing and thawing some phase separation might occur - Please ensure that the solution is mixed thoroughly but GENTLY before use.

    This antibody (ab6556) is the purifed version of our best-selling rabbit polyclonal to GFP (ab290). It has been developed specifically for use in applications requiring a high titre and specificity with minimum background such as immuno-electron microscopy.

    This anti-GFP antibody recognizes the enhanced form of GFP as well.

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 or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液
    pH: 7.40
    Constituents: 0.79% Tris HCl, 25% Glycerol
  • Concentration information loading...
  • 纯度
    Immunogen affinity purified
  • 纯化说明
    This antibody is an affinity purified rabbit anti-GFP antibody purified on an affinity chromatography column made with highly purified recombinant GFP.
  • 克隆
    多克隆
  • 同种型
    IgG
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab6556 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-FoFr Use at an assay dependent concentration. PubMed: 19563657
Electron Microscopy 1/5000.
IHC-P 1/1000. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
IHC-Fr 1/1000.
IP Use at an assay dependent concentration. Use Protein A agarose for IP.
WB 1/1000 - 1/5000.
ICC/IF 1/2000.
Flow Cyt Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

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

  • Regionally restricted CX3CL1 expression in the retina of adult CX3CL1 cherry: CX3CR1gfp mouse (A-E)

    Fluorescent light microscopy analysis of a vertical retina cryosection (A) showing both CX3CL1/Cherry (detected in rhodamine channel, red) (B) and CX3CR1/GFP (detected in FITC channel, green) (C) reporters expression. CX3CR1/GFP signal is assigned to microglial cells with cell bodies and dendritic ramifications in three sublayers (outer plexiform layer, OPL, inner plexiform layer, IPL and ganglion cells layer, GCL). CX3CL1/Cherry fluorescent cells are found exclusively in ganglion cell and inner nuclear layer. No fluorescent cells were observed in outer nuclear layer (ONL). Enlargements show strong Cherry signal in a presumable amacrine cell body in inner nuclear (D, arrow points to dendritic base) and ganglion cell (E) layers, indicating high level of the CX3CL1 chemokine promotor activity in neurons. Cryosections are counterstained with DAPI nucleic acid stain (blue). Distribution of the Cherry reporter positive cells in the retina of CX3CL1cherry: CX3CR1gfp mice coincides mostly with results obtained in in situ hybridization studies. Of note, membrane/lipid inner and outer photoreceptor segment layer exhibit nonspecific fluorescence. Scale bars represent 50 µm (A, B and C), and 10 µm (D, E).

    GFP was detected with ab6556 at 1/1000 dilution.

    (From Figure 4 of Zieger et al)

  • The downstream Wnt non-canonical pathway components are required in the escort cells for cystoblast encapsulation

    (A-D1) Germaria of c587-GAL4, RhoA, Rac1 and cdc42 depleted escort cells stained with Tj (red), GFP (green) and Vasa (blue) showing loss of encapsulation in RhoA, Rac1 and cdc42 depleted escort cells. Fax-GFP marks somatic cell membranes. GFP channel is shown in A1, B1, C1 and D1. Scale bar for all images is 20μm.

    GFP is detected with ab6556.

    (After Figure 2 of Upadhyay et al)

  • All lanes : Anti-GFP antibody (ab6556) at 1/5000 dilution

    Lane 1 : LNCaP cells pEGFP-PKD1 empty vector
    Lane 2 : LNCaP pEGFP-PKD1 transfected cells

    Secondary
    All lanes : Rabbit polyclonal to GFP (ab6556) at 1/10000 dilution (Goat anti-rabbit HRP)

    Developed using the ECL technique.

    Performed under reducing conditions.

    Exposure time: 10 seconds

    See Abreview

  • ab6556 staining GFP in mouse tooth tissue section by Immunohistochemistry (Frozen sections) without fixation. Tissue samples were blocked with 1% BSA for 20 minutes at 20oC. The sample was incubated with primary antibody (1/500) for 2 hours at 20oC. An Alexa Fluor®488-conjugated Chicken polyclonal to rabbit IgG was used as secondary antibody at 1/200 dilution. Immunofluorescent localization of CD31 and GFP in implanted non GFP cultured molars in GFP adult mice showed the origin of neo-formed blood vessels.
    DP: Dental Pulp
    EO: Enamel Organ

    See Abreview

  • This image shows a single primary hippocampal neuron from a primary culture overexpressing GFP stained with ab6556 at a dilution of 1/2000. This picture was kindly supplied as part of the review submitted by one of our customers.

  • This image shows IF using GFP-expressing glial cells (green) transplanted into lesioned rat spinal cord. This was detected using ab6556 anti-GFP antibody and a FITC conjugated secondary antibody. Axons are labelled red by an antibody to neurofilament-200 and a rhodamine secondary. ab6556 reveals the morphology of the transplanted cells to such an extent that their close interactions with axons are obvious. The top picture shows an optical section from a confocal microscope scan showing how a GFP cell wraps around a branched axon travelling longitudinally. The bottom picture consists of an optical section from another confocal scan showing a GFP cell enveloping an axon in the transverse plane. Review by Andrew Toft submitted 19 May 2004.

  • ab6556 at 1/500 dilution staining GFP in mouse testis by immunohistochemistry (frozen sections). Sections were paraformaldehyde fixed prior to blocking in 100% serum for 1 hour at 37°C and then incubated with ab6556 for 1 hour at 37°C. A Texas Red conjugated chicken polyclonal to rabbit Ig, diluted 1/100, was used as the secondary antibody.

    See Abreview

  • Specific labeling of a Trk-GFP fusion protein being synthesized on ER in sympathetic neurons infected with an adenovirus carrying the construct. The gold is associated with the ER membranes. This was done using a 1/5000 dilution of affinity purified antibody (ab6556). The tissue section was fixed and embedded using durcupan resin.

文献

This product has been referenced in:
See all 578 Publications for this product

客户评价及客户问答

1-10 of 77 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK 293 cells)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
30 µg
Specification
HEK 293 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

提交于 Apr 23 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Sheep Cell (lung)
Permeabilization
No
Specification
lung
Blocking step
Blockaid as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

提交于 May 31 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (fat)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mm sodium citrate PH6.0
Permeabilization
No
Specification
fat
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Paraformaldehyde

Dr. Jiang Hu

Verified customer

提交于 Mar 16 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Zebrafish Cell (Wholemount Embryo)
Permeabilization
Yes - 1% DMSO + 0.5% Triton X
Specification
Wholemount Embryo
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

提交于 Jan 03 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Lung carcinoma cells transfected with CMV-GFP, xen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: NaCitrate pH 6
Permeabilization
No
Specification
Lung carcinoma cells transfected with CMV-GFP, xen
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: 20°C
Fixative
Formaldehyde

Abcam user community

Verified customer

提交于 May 17 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Mouse ES cell (YFP) reporter line.)
Permeabilization
Yes - 5% donkey serum in PBST (PBS / 0.1% v/v Triton X-100 (Sigma))
Specification
Mouse ES cell (YFP) reporter line.
Blocking step
5% donkey serum in PBST (PBS / 0.1% v/v Triton X-100 (Sigma)) as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

提交于 Sep 21 2015

Application
Immunocytochemistry
Sample
Human Cell (HEK293T overexpressing GFP)
Permeabilization
Yes - 0.1% TRITONX1000.
Specification
HEK293T overexpressing GFP
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

提交于 May 12 2015

Application
Western blot
Loading amount
400000 cells
Gel Running Conditions
Reduced Denaturing (12%)
Sample
Mouse Cell lysate - whole cell (Mouse Neuroblastoma (Neuro2A))
Specification
Mouse Neuroblastoma (Neuro2A)
Treatment
10% FBS and fibroblast growth factor
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 28°C

Abcam user community

Verified customer

提交于 Apr 01 2015

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (LNCaP cells)
Specification
LNCaP cells
Treatment
pEGFP-PKD1 transfected
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

提交于 Feb 13 2015

1-10 of 77 Abreviews or Q&A

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

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