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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: IP, 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.
  • 经测试应用

    适用于: IP, ICC/IF, WBmore details
  • 种属反应性

    与反应: Species independent
  • 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.
  • 常规说明

    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. 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

Abpromise™承诺保证使用ab291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 (2)
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
IP
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

  • SDS download

  • Datasheet download

    Download

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

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
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 Mar 20 2014

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
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.

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
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.

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
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.

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
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. 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
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. Leszek Lisowski

Verified customer

提交于 Jan 06 2009

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

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

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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