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Tags & Cell Markers Fusion / Marker Proteins GFP

生物素Anti-GFP抗体(ab6658)

  • Datasheet
  • SDS
Reviews (6)Q&A (12)References (115)

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Western blot - Biotin Anti-GFP antibody (ab6658)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-GFP antibody (ab6658)
  • IHC - Wholemount - Biotin Anti-GFP antibody (ab6658)
  • Immunohistochemistry (Frozen sections) - Biotin Anti-GFP antibody (ab6658)
  • Immunocytochemistry/ Immunofluorescence - Biotin Anti-GFP antibody (ab6658)

Key features and details

  • Biotin Goat polyclonal to GFP
  • Suitable for: WB, IP, ICC/IF, Sandwich ELISA, IHC-P, IHC-Fr
  • Reacts with: Species independent
  • Conjugation: Biotin
  • Isotype: IgG

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概述

  • 产品名称

    生物素Anti-GFP抗体
    参阅全部 GFP 一抗
  • 描述

    生物素山羊多克隆抗体to GFP
  • 宿主

    Goat
  • 偶联物

    Biotin
  • 特异性

    Antibody recognizes wild type, recombinant and enhanced forms of GFP (EGFP). No reaction was observed against Human, Mouse and Rat Serum Proteins.
  • 经测试应用

    适用于: WB, IP, ICC/IF, Sandwich ELISA, IHC-P, IHC-Frmore details
  • 种属反应性

    与反应: Species independent
  • 免疫原

    Recombinant full length protein corresponding to GFP aa 1-246.
    Sequence:

    MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTT GKLPVPWPTL VTTFSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTI FFKDDGNYKTRAEVKFEGDTLV NRIELKGIDFKEDGNILGHKLEYNYN SHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLAD HYQQNTPIGDGPVL LPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK


    Database link: P42212
    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 常规说明

    Designed to detect GFP and its variants in immunoblotting and immunoprecipitation.

    Biotinamidocaproate N-Hydroxysuccinimide Ester (BAC) Biotin/Protein Ratio: 10-20 BAC molecules per goat IgG molecule.

    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. Avoid freeze / thaw cycle.
  • 存储溶液

    Preservative: 0.01% Sodium azide
    Constituents: 0.44% Potassium phosphate, 0.88% Sodium chloride

    10 mg/mL BSA, immunoglobulin and protease free
  • Concentration information loading...
  • 纯度

    Affinity purified
  • 纯化说明

    This product was prepared from monospecific antiserum by immunoaffinity chromatography using Green Fluorescent Protein (Aequorea victoria) coupled to agarose beads followed by solid phase adsorption(s) to remove any unwanted reactivities.
  • Primary antibody说明

    Designed to detect GFP and its variants in ELISA (sandwich or capture), immunoblotting and immunoprecipitation.
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

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

相关产品

  • Isotype control

    • Biotin Goat IgG - Isotype Control (ab37376)
  • Recombinant Protein

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

    • GFP ELISA Kit (ab171581)
    • Biotin Assay Kit (Colorimetric) (ab185441)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab6658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
WB (2)
1/2000 - 1/10000.
IP (1)
Use at an assay dependent concentration.
ICC/IF (1)
1/1000 - 1/5000.
Sandwich ELISA
Use at an assay dependent concentration.
IHC-P (2)
1/250.
IHC-Fr
1/5000.
说明
WB
1/2000 - 1/10000.
IP
Use at an assay dependent concentration.
ICC/IF
1/1000 - 1/5000.
Sandwich ELISA
Use at an assay dependent concentration.
IHC-P
1/250.
IHC-Fr
1/5000.

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

  • Western blot - Biotin Anti-GFP antibody (ab6658)
    Western blot - Biotin Anti-GFP antibody (ab6658)
    Biotin Anti-GFP antibody (ab6658)
    Additional bands at: 28 kDa. We are unsure as to the identity of these extra bands.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-GFP antibody (ab6658)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Biotin Anti-GFP antibody (ab6658)This image is a courtesy of Hongwei Shao

    ab6658 staining GFP in human melanoma cells recovered from nude mice by Immunocytochemistry/ immunoflurescence. Cells were fixed with formaldehyde, permeabilized with 0.25% Triton X-100 RT for 10min and blocking with commercially available blocking buffer was performed for 30 minutes at RT. Samples were incubated with primary antibody (1:50) for 18 hours at 4°C. An Alexa Fluor®488-conjugated donkey polyclonal to goat IgG was used as secondary antibody at 1/100 dilution. Green color indicates GFP/Fibrolast cells, while red color indicates Ki67 positive cells, most of them are tumor cells (Abcam`s ab15580 was used for the detection).

    See Abreview

  • IHC - Wholemount - Biotin Anti-GFP antibody (ab6658)
    IHC - Wholemount - Biotin Anti-GFP antibody (ab6658)

    Immunofluorescence Microscopy using ab6658. Tissue: Drosophila melanogaster late stage embryonic central nervous system. Fixation: 0.5% PFA. Antigen retrieval: not required. Primary antibody: Anti-GFP antibody at a 1/1,000 for 1 h at RT. Secondary antibody: AlexaFluor 488™ conjugated anti-Goat antibody at 1/300 for 45 min at RT. Panel A: shows a lateral view (ventral left). Panels B and C: shows ventral views of whole mount embryos at 63x magnification (plus 2x digital zoom). In all panels, anterior is up. Staining: tau-GFP cell bodies (large arrowhead) and axons of motorneurons (arrow) and interneurons (small arrowhead) as green fluorescent signal.

  • Immunohistochemistry (Frozen sections) - Biotin Anti-GFP antibody (ab6658)
    Immunohistochemistry (Frozen sections) - Biotin Anti-GFP antibody (ab6658)

    Immunofluorescence Microscopy using ab6658. Tissue: Sf-1:Cre mice crossed to the Z/EG reporter line. Mouse brain (coronal view, 20X magnification). Fixation: 4%PFA/PBS with o/n fixation, and subsequently transferred to a 30% sucrose solution. Antigen retrieval: frozen in OCT freezing medium (Sakura) and cryostat sectioned at 40 microns. Primary antibody: Goat anti- GFP was used at 1/500 dilution in free floating imunnohistochemistry to detect GFP. Secondary antibody: Fluorchrome conjugated Anti-goat IgG secondary antibody was used for detection at 1:10,000 for 45 min at RT.Localization: Sf-1+ neurons and their processes of the ventromedial nucleus of the hypothalamus. Staining: eGFP as green fluorescent signal and sections were counterstained with DAPI.

  • Immunocytochemistry/ Immunofluorescence - Biotin Anti-GFP antibody (ab6658)
    Immunocytochemistry/ Immunofluorescence - Biotin Anti-GFP antibody (ab6658)Image courtesy of Efrat Shema by Abreview.

    ab6658 staining GFP in rat brain cells infected with viruses containing GFP under a CMV promoter by Immunocytochemistry/ Immunofluorescence.Cells were fixed with formaldehyde, permeabilized using 0.2% Triton, blocked with 20% serum and then incubated with ab6658 at a 1:50 dilution for 20 hours at 25°C. The secondary used was an Alexa-Fluor 488 conjugated rabbit polyclonal, used at a 1/200 dilution.

    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 (115)

发表研究结果有使用 ab6658?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab6658 被引用在 115 文献中.

  • He J  et al. DNA methylation maintenance at the p53 locus initiates biliary-mediated liver regeneration. NPJ Regen Med 7:21 (2022). PubMed: 35351894
  • Schamber MR & Vafabakhsh R Mechanism of sensitivity modulation in the calcium-sensing receptor via electrostatic tuning. Nat Commun 13:2194 (2022). PubMed: 35459864
  • Niu L  et al. Negative Regulator Nlrc3-like Maintain the Balanced Innate Immune Response During Mycobacterial Infection in Zebrafish. Front Immunol 13:893611 (2022). PubMed: 35693809
  • Fung CW  et al. Cell fate determining molecular switches and signaling pathways in Pax7-expressing somitic mesoderm. Cell Discov 8:61 (2022). PubMed: 35764624
  • Li X  et al. Estrogens revert neutrophil hyperplasia by inhibiting Hif1α-cMyb pathway in zebrafish myelodysplastic syndromes models. Cell Death Discov 8:323 (2022). PubMed: 35842445
View all Publications for this product

客户评价及客户问答

Show All 评价 Q&A
提交评价 提交问题

1-10 of 18 Abreviews or Q&A

Immunoprecipitation abreview for Anti-GFP antibody (Biotin)

Good
Abreviews
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (E. coli)
Total protein in input
4000 µg
Immuno-precipitation step
Other - Dynabeads
Specification
E. coli
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Feb 26 2018

Western blot abreview for Anti-GFP antibody (Biotin)

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Fibroblast)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
30 µg
Specification
Fibroblast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Hongwei Shao

Verified customer

提交于 Sep 06 2017

Immunocytochemistry/ Immunofluorescence abreview for Anti-GFP antibody (Biotin)

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Brain tissue slices)
Permeabilization
Yes - Triton 0.2%
Specification
Brain tissue slices
Blocking step
Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 20% · Temperature: 25°C
Fixative
Formaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Efrat Shema

Verified customer

提交于 Aug 04 2011

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-GFP antibody (Biotin)

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (Brain)
Specification
Brain
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Sodium citrate pH 6.0
Permeabilization
Yes - Triton 0.2%
Blocking step
Serum as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 20%
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Dr. Efrat Shema

Verified customer

提交于 Aug 02 2011

Question

Hello,
Is it possible to have an estimate of the binding constant (kON - kOFF) of the following product:
ab6658

Read More

Abcam community

Verified customer

Asked on Feb 10 2015

Answer



I am sorry to confirm that the binding constant is not often determined for research grade antibodies and there is no binding constant information available for this particular product.

I am sorry we have no data to share to help answer your question on this occasion.

Read More

Sam Washer

Abcam Scientific Support

回复于 Feb 10 2015

Question

What is the molecular weight of this biotinylated antibody?

Read More

Abcam community

Verified customer

Asked on Aug 13 2014

Answer


I performed the following molecular weight calculations for this antibody:

Assuming 15 biotin molecules per IgG molecule:

MW IgG = 150 kDa
MW biotin = 244.31 g/mol = 244.31 Da = 0.24431 kDa
MW ab6658 = 150 + (15*0.24431) ˜ 154 kDa

So the molecular weight of the biotinylated antibody should be ˜ 154 kDa.

Read More

Kevin Hanson

Abcam Scientific Support

回复于 Aug 13 2014

Question

What percentage is conjugated to biotin?

Read More

Abcam community

Verified customer

Asked on Nov 01 2012

Answer

We do not determine the percentage of molecules conjugated to biotin, but it is expected to be close to 100%. The conjugation ratio between our biotin and IgG is usually 5-6 molecules of biotin per molecule of IgG.

Read More

Abcam Scientific Support

回复于 Nov 01 2012

Question


What is the unconjugated equivalent to Anti-GFP antibody (Biotin) (ab6658)?

Read More

Abcam community

Verified customer

Asked on Sep 28 2012

Answer

The unconjugated version of ab6658 is ab6673.

https://www.abcam.com/GFP-antibody-ab6673.html

Read More

Abcam Scientific Support

回复于 Sep 28 2012

Question

Inquiry: I am using your biotin-conjugated anti-GFP (ab6658). How does the signal from this antibody compare to one that is not biotin-conjugated and requires a biotin-conjugated secondary antibody? Is the signal equal to, less than, or greater than using an unconjugated primary with a secondary? I am using ABC reagent to amplify the signal. Thanks!

Read More

Abcam community

Verified customer

Asked on Aug 30 2012

Answer

Thank you very much for contacting us with your question.

In general, using a secondary antibody will help amplify the signal from the primary antibody. With a pre-conjugated primary antibody, the signal may be weaker than using a conjugated secondary antibody. The avidin-biotin complex will also help amplify the signal, and there are other benefits to using a pre-conjugated primary (no cross-reaction between the secondary and samples, faster assay, etc), so all of these points should be weighed when considering whether to use a pre-conjugated primary or a secondary antibody step.

Read More

Abcam Scientific Support

回复于 Aug 30 2012

Question

I have a question regarding the ab6658 primary antibody. Currently, we are using it for detection of our mouse tissue.
I would like to start using it for the detection of rat GFP now, is it possible to use this antibody or do you recommend a better one?
If possible, I would prefer the DAB detection above the fluorescent one.
Thanks.
Best regards,

Read More

Abcam community

Verified customer

Asked on Jul 09 2012

Answer

>pecies specificity is not applicable for this product. You can try this antibody for detection of GFP in any type of cells if they express GFP protein.

Read More

Abcam Scientific Support

回复于 Jul 09 2012

1-10 of 18 Abreviews or Q&A

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