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Tags & Cell Markers Fusion / Marker Proteins GFP

Anti-GFP抗体(ab5450)

  • Datasheet
  • SDS
Reviews (11)Q&A (6)References (158)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab5450)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab5450)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab5450)
  • IHC - Wholemount - Anti-GFP antibody (ab5450)
  • Electron Microscopy - Anti-GFP antibody (ab5450)

Key features and details

  • Goat polyclonal to GFP
  • Suitable for: IP, ICC, IHC-FoFr, Electron Microscopy, IHC - Wholemount, ICC/IF, WB, IHC-P
  • Reacts with: Species independent
  • Isotype: IgG

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概述

  • 产品名称

    Anti-GFP抗体
    参阅全部 GFP 一抗
  • 描述

    山羊多克隆抗体to GFP
  • 宿主

    Goat
  • 经测试应用

    适用于: IP, ICC, IHC-FoFr, Electron Microscopy, IHC - Wholemount, ICC/IF, WB, IHC-Pmore details
  • 种属反应性

    与反应: Species independent
  • 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

    Protein A will not bind goat IgG, so use alternates (eg. protein G) in IP with this antibody.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

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

    Protein G purified
  • 纯化说明

    This antibody is Goat anti-GFP serum (ab5449) affinity purified using a HiTrap-NHS activated sepharose column (Amersham-Pharmacia) containing covalently linked highly purified recombinant GFP. After applying the serum to the column and extensive washing, the eluted anti-GFP immunoglobulins are desalted.
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Tags & Cell Markers
    • Fusion / Marker Proteins
    • GFP

相关产品

  • Compatible Secondaries

    • Donkey Anti-Goat IgG H&L (Alexa Fluor® 488) (ab150129)
    • Donkey Anti-Goat IgG H&L (HRP) (ab205723)
  • Isotype control

    • Goat IgG, polyclonal - Isotype Control (ab37373)
  • Recombinant Protein

    • Recombinant E. coli GFP protein (His tag) (ab119740)
    • Recombinant A. victoria GFP protein (ab84191)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
    • GFP ELISA Kit (ab171581)
    • Anti-GFP antibody (ab5449)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab5450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
IP (1)
Use at an assay dependent concentration.
ICC
Use at an assay dependent concentration.
IHC-FoFr
Use at an assay dependent concentration.
Electron Microscopy
Use at an assay dependent concentration.
IHC - Wholemount (2)
Use at an assay dependent concentration.
ICC/IF (2)
1/2000.
WB
1/2000 - 1/20000.
IHC-P (4)
1/1000.
说明
IP
Use at an assay dependent concentration.
ICC
Use at an assay dependent concentration.
IHC-FoFr
Use at an assay dependent concentration.
Electron Microscopy
Use at an assay dependent concentration.
IHC - Wholemount
Use at an assay dependent concentration.
ICC/IF
1/2000.
WB
1/2000 - 1/20000.
IHC-P
1/1000.

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) - Anti-GFP antibody (ab5450)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab5450)Image from Sylva M et al., Plos One. 2011;6(10):e26088. Fig 2.; doi: 10.1371/journal.pone.0022616 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    Immunohistochemical staining of donor BMDCs expressing GFP in mouse pancreas tissue using ab5450. Antigen retrieval was carried out using target-retrieval solution for 30 minutes in a boiling water bath. Endogenous peroxidase activity was quenched with 3% hydrogen peroxide (5 minutes). Panels A and B were detected with a polymer horseradish peroxidase anti-rabbit detection system (30 minutes) and 3,3′-diaminobenzidine was used as a substrate. Counter-staining was performed with hematoxylin. 

  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab5450)
    Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody (ab5450)

    ab5450 staining GFP in GFP-transfected NIH3T3 cells. The cells were fixed with 4% formaldehyde (10min) and then blocked in 1% BSA / 0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab5450 at 1/2000 dilution overnight at +4°C followed by incubation with ab150129, Donkey Anti-Goat IgG H&L (Alexa Fluor® 488), for 1 hour, at 1μg/ml.
    Under identical experimental conditions, when compared to the basal level of GFP expression in transfected NIH3T3 cells, the cells upon which ab5450 was applied gave a stronger signal in the 488 channel, indicating that ab5450 is binding to GFP and therefore eliciting signal amplification.
    ab5450 was also applied to non-GFP-transfected NIH3T3 cells, which produced no positive staining, indicating specificity for GFP. Nuclear DNA was labelled with 1.43μM DAPI (blue).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab5450)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GFP antibody (ab5450)Image courtesy of an anonymous Abreview.
    ab5450 staining GFP in murine brain tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with paraformaldehyde, permeabilized using 0.2% Triton and then blocked with serum for 1 hour at room temperature, followed by incubation with the primary antibody at a 1/1000 dilution for 14 hours at 4°C. A Cy2®-conjugated secondary antibody was used at a 1/200 dilution.
  • IHC - Wholemount - Anti-GFP antibody (ab5450)
    IHC - Wholemount - Anti-GFP antibody (ab5450)Image courtesy of Sana Zakaria by Abreview.
    ab5450 staining GFP in murine hindbrain tissue by IHC-Wholemount.
    Dissected hindbrains of E12.5 stage embryos were fixed in 4% paraformaldehyde for 4-5 hours and then washed in PBS before proceeding with immunolabelling in PBS with 1% triton. Primary antibody was incubated for 4-6 days (99-144 hours). The hindbrain was cultured as an explant after GFP was electroporated. The wholemount immuno with ab5450 at a 1/500 dilution was done to see if GFP could be detected, it was the same GFP expression with the antibody as that which had been seen initially with the electroporation. The secondary used was an Alexa-Fluor 647 conjugated donkey anti-goat polyclonal used at a 1/150 dilution. Blue is DAPI and green is the GFP antibody (pseudo-colored from red Alexa Fluor 647).

    See Abreview

  • Electron Microscopy - Anti-GFP antibody (ab5450)
    Electron Microscopy - Anti-GFP antibody (ab5450)This image is courtesy of an Abreview submitted by Narciso Campos

    Electron Microscopy of Arabidopsis thaliana tissue sections labelling GFP with ab5450. An 18nm gold-conjugated Donkey anti-goat IgG polyclonal (1/15) was used as the secondary antibody.

     

    Arabidopsis thaliana transgenic plant expressing GFP fused to an Endoplasmic Recticulum (ER) marker. No label is observed in: C - chloroplasts, M - mitochondria, G - golgi. The sample was prepared by cryofixation and embedding in Lowicryl HM20 resin. The image was taken in a JEOL 1010 transmission electron microscope at 20,000× magnification.

    See Abreview

实验方案

  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (158)

发表研究结果有使用 ab5450?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab5450 被引用在 158 文献中.

  • Zhou YQ  et al. Genetically engineered distal airway stem cell transplantation protects mice from pulmonary infection. EMBO Mol Med 12:e10233 (2020). PubMed: 31782624
  • Diao L  et al. Increased proliferation of hepatic periportal ductal progenitor cells contributes to persistent hypermetabolism after trauma. J Cell Mol Med 24:1578-1587 (2020). PubMed: 31793707
  • Costamagna D  et al. Interleukin-4 administration improves muscle function, adult myogenesis, and lifespan of colon carcinoma-bearing mice. J Cachexia Sarcopenia Muscle 11:783-801 (2020). PubMed: 32103619
  • Chillappagari S  et al. Defective BACH1/HO-1 regulatory circuits in cystic fibrosis bronchial epithelial cells. J Cyst Fibros N/A:N/A (2020). PubMed: 32534959
  • Eagle AL  et al. Circuit-specific hippocampal ?FosB underlies resilience to stress-induced social avoidance. Nat Commun 11:4484 (2020). PubMed: 32901027
View all Publications for this product

客户评价及客户问答

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1-2 of 2 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-GFP antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Testis)
Permeabilization
No
Specification
Testis
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 24°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

提交于 Apr 14 2021

Immunocytochemistry/ Immunofluorescence abreview for Anti-GFP antibody

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Brain, cortical neurons)
Specification
Brain, cortical neurons
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% Triton X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Read More

Abcam user community

Verified customer

提交于 Oct 27 2009

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