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Tags & Cell Markers Fusion / Marker Proteins GFP

Anti-GFP抗体[LGB-1] (ab291)

  • Datasheet
  • SDS
Reviews (6)Q&A (5)References (24)

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Western blot - Anti-GFP antibody [LGB-1] (ab291)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [LGB-1] (ab291)
  • Immunoprecipitation - Anti-GFP antibody [LGB-1] (ab291)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [LGB-1] (ab291)
  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [LGB-1] (ab291)

Key features and details

  • Mouse monoclonal [LGB-1] to GFP
  • Suitable for: Flow Cyt, IP, ELISA, ICC/IF, WB
  • Reacts with: Species independent
  • Isotype: IgG1

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概述

  • 产品名称

    Anti-GFP抗体[LGB-1]
    参阅全部 GFP 一抗
  • 描述

    小鼠单克隆抗体[LGB-1] to GFP
  • 宿主

    Mouse
  • 特异性

    This antibody recognizes all forms of GFP from Aquorea victoria (i.e. GFP, EGFP, YFP and CFP). See Abreview for CFP immunoprecipitation.
  • Tested Applications & Species

    Application Species
    ICC/IF
    Species independent
    See all applications and species data
  • 免疫原

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

  • 阳性对照

    • Pure GFP protein, or cells known to overexpress GFP.

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • 存储溶液

    pH: 7.20
    Constituents: PBS, 50% Glycerol
  • Concentration information loading...
  • 纯度

    Protein A purified
  • 克隆

    单克隆
  • 克隆编号

    LGB-1
  • 同种型

    IgG1
  • 轻链类型

    kappa
  • 研究领域

    • Tags & Cell Markers
    • Fusion / Marker Proteins
    • GFP
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • Zinc Finger
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • Nuclear Hormone Receptors
    • Estrogen

相关产品

  • Alternative Versions

    • Biotin Anti-GFP antibody [LGB-1] (ab42562)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
  • Recombinant Protein

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

    • GFP ELISA Kit (ab171581)

应用

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab291 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

应用 Species
ICC/IF
Species independent
应用 Ab评论 说明
Flow Cyt
Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

IP (2)
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.
ICC/IF (1)
Use a concentration of 0.5 µg/ml.
WB (2)
Use a concentration of 0.5 µg/ml. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).
说明
Flow Cyt
Use at an assay dependent concentration.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

 

IP
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.
ICC/IF
Use a concentration of 0.5 µg/ml.
WB
Use a concentration of 0.5 µg/ml. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).

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 - Anti-GFP antibody [LGB-1] (ab291)
    Western blot - Anti-GFP antibody [LGB-1] (ab291)
    All lanes : Anti-GFP antibody [LGB-1] (ab291) at 0.5 µg/ml

    Lane 1 : 5ng GFP
    Lane 2 : 10ng GFP
    Lane 3 : 25ng GFP

    Secondary
    All lanes : Sheep anti-mouse IgG HRP conjugate at 1/5000 dilution

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

  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [LGB-1] (ab291)
    Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [LGB-1] (ab291)
    Paraformaldehyde fixed COS-7 cells expressing Myr-N15-PAK2-EGFP construct (Vilas et al.(2006) PNAS 103, 6542). Myr-N15-PAK2-EGFP fluorescence is shown in green. Indirect immunofluorescenct detection of N15-PAK-EGFP using ab291 monoclonal LGB-1 anti-GFP at 0.05 ug/ml with chicken anti-mouse secondary antibody conjugated to Alexa594 diluted 1/500 is shown in red. Myr-N15-PAK2-EGFP is localized to membrane ruffles and perinuclear vesicular structures (likely Golgi,TGN or late endosomes).
  • Immunoprecipitation - Anti-GFP antibody [LGB-1] (ab291)
    Immunoprecipitation - Anti-GFP antibody [LGB-1] (ab291)This image was kindly supplied by Dr Lindsay Tulloch by Abreview
    ab291 at 6.7µg/mg lysate.
    HEK293 Cell lysate at 300µg.
    Transfected with CFP-fused protein XXX in pECFP vector. Immunoprecipitation step using Protein G.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [LGB-1] (ab291)
    Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [LGB-1] (ab291)This image is courtesy of an Abreview submitted by Vladimir Milenkovic
    ab291 staining GFP in Dog MDCKII cells transfected with GFP by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% TX100 and blocked with 5% serum for 20 minutes. Samples were incubated with primary antibody (1/250 PBS + 0.1% TX100 + 1% goat serum) for 16 hours at 4°C. An Alexa Fluor®546-conjugated Goat anti-mouse IgG polyclonal (1/500) was used as the secondary antibody. DAPI was used to stain nuclei. ab291 was used to assess electoporation efficiency of double transfected MDCKII cells.

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [LGB-1] (ab291)
    Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [LGB-1] (ab291)

    ab291 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 ab291 at 1/200 dilution overnight at +4°C followed by incubation with ab150117, Goat Anti-Mouse 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 ab291 was applied gave a stronger signal in the 488 channel, indicating that ab291 is binding to GFP and therefore eliciting signal amplification.
    ab291 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).

实验方案

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

    • Datasheet
    • SDS
  • 文献 (24)

    发表研究结果有使用 ab291?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab291 被引用在 24 文献中.

    • Griffin JM  et al. Astrocyte-selective AAV gene therapy through the endogenous GFAP promoter results in robust transduction in the rat spinal cord following injury. Gene Ther 26:198-210 (2019). PubMed: 30962538
    • Bigot N  et al. Phosphorylation-mediated interactions with TOPBP1 couple 53BP1 and 9-1-1 to control the G1 DNA damage checkpoint. Elife 8:N/A (2019). PubMed: 31135337
    • Giesecke T  et al. Vasopressin Increases Urinary Acidification via V1a Receptors in Collecting Duct Intercalated Cells. J Am Soc Nephrol 30:946-961 (2019). PubMed: 31097611
    • Lyytinen OL  et al. Microbial production of lipid-protein vesicles using enveloped bacteriophage phi6. Microb Cell Fact 18:29 (2019). PubMed: 30732607
    • Zhang L  et al. A GABAergic cell type in the lateral habenula links hypothalamic homeostatic and midbrain motivation circuits with sex steroid signaling. Transl Psychiatry 8:50 (2018). PubMed: 29479060
    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 11 Abreviews or Q&A

    Question

    Will this antibody react with the mutant GFP-cycle3 (from Invitrogen, TopoGlo plasmid) ?

    Read More

    Abcam community

    Verified customer

    Asked on Oct 01 2015

    Answer

    mutant GFP-cycle3 has not been tested with this antibody

    Read More

    Heather Allen

    Abcam Scientific Support

    回复于 Oct 01 2015

    Immunohistochemistry (Frozen sections) abreview for Anti-GFP antibody [LGB-1]

    Poor
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    提交于 Mar 20 2014

    Question

    Will this antibody detect in the picomol-nanomol range in ELISA?

    Read More

    Abcam community

    Verified customer

    Asked on Jan 19 2011

    Answer

    Ab291 should be detecting nanomolar amounts of antigen, and very likely picomolar as well in ELISA. In Western blotting it readily detects 5ng GFP (0.2 picomoles) in a film exposure of only a few seconds.

    Read More

    Abcam Scientific Support

    回复于 Jan 19 2011

    Immunoprecipitation abreview for Anti-GFP antibody [LGB-1]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - whole cell (HEK 293 cells transfected with Rab27a-GFP)
    Total protein in input
    25 µg
    Specification
    HEK 293 cells transfected with Rab27a-GFP
    Immuno-precipitation step
    Protein G
    Read More

    Herr Dr. Vladimir Milenkovic

    Verified customer

    提交于 Jul 22 2010

    Western blot abreview for Anti-GFP antibody [LGB-1]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HEK 293 cells transfected with Rab27a-GFP)
    Loading amount
    25 µg
    Specification
    HEK 293 cells transfected with Rab27a-GFP
    Gel Running Conditions
    Reduced Denaturing (10%)
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Herr Dr. Vladimir Milenkovic

    Verified customer

    提交于 Jul 22 2010

    Immunocytochemistry/ Immunofluorescence abreview for Anti-GFP antibody [LGB-1]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Dog Cell (MDCK II cells transfected with GFP)
    Specification
    MDCK II cells transfected with GFP
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.5% TX100
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 5%
    Read More

    Herr Dr. Vladimir Milenkovic

    Verified customer

    提交于 Jun 16 2010

    Immunoprecipitation abreview for Anti-GFP antibody [LGB-1]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunoprecipitation
    Sample
    Human Cell lysate - other (HEK 293)
    Total protein in input
    300 µg
    Specification
    HEK 293
    Treatment
    Transfected with CFP-fused protein XXX in pECFP vector
    Immuno-precipitation step
    Protein G
    Read More

    Dr. Lindsay Tulloch

    Verified customer

    提交于 Dec 10 2009

    Western blot abreview for Anti-GFP antibody [LGB-1]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Hela, Huh7 and 293 cells)
    Loading amount
    10 µg
    Specification
    Hela, Huh7 and 293 cells
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis-Tris Gel)
    Blocking step
    Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Read More

    Dr. Leszek Lisowski

    Verified customer

    提交于 Jan 06 2009

    Question

    Is this anti-GFP antibody or any other anti-GFP antibodies raised by using adjuvant which does not contain Mycobacterial components? As you know, common adjuvant such as Freund's complete adjuvant contains mycobacterial cell extracts. I worry this because I want to detect GFP expressed in mycobacteria, and it is essential to use antibodies with no cross reactivity to mycobacterial components.

    Read More

    Abcam community

    Verified customer

    Asked on Jun 02 2008

    Answer

    Adjuvant was used with this ab290 and may not be suitable for your research. However, we have a few other products that you may be interested in. 1) ab291 GFP antibody [LGB-1]: No adjuvant was used 2) ab5450 GFP antibody: The affinity purification on a GFP column should have removed all interfering antibodies 3) ab6556 GFP antibody: The affinity purification on a GFP column should have removed all interfering antibodies

    Read More

    Abcam Scientific Support

    回复于 Jun 02 2008

    Question

    resp. sir can we use the same antibody for staining mammalian brain sections(including human), and that too on perfusion fixed(excluding human) cryosections using PAP protocol

    Read More

    Abcam community

    Verified customer

    Asked on Jun 23 2001

    Answer

    We have been unable to find any information on whether the estrogen receptor is expressed in brain - this antibody was primarily produced for the diagnosis of breast tumours. Secondly, although this antibody can be used in frozen (cryostat) sections, we do not know whether it recognises the antigen in species other than human. Nevertheless, the PAP staining method should be fine.

    Read More

    Neil Hayward

    Abcam Scientific Support

    回复于 Jun 25 2001

    1-10 of 11 Abreviews or Q&A

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