Anti-GFP抗体[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
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 -
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
相关产品
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
Recombinant Protein
-
Related Products
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
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
-
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 -
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)This image was kindly supplied by Dr Lindsay Tulloch by Abreviewab291 at 6.7µg/mg lysate.
HEK293 Cell lysate at 300µg.Transfected with CFP-fused protein XXX in pECFP vector. Immunoprecipitation step using Protein G. -
Immunocytochemistry/ Immunofluorescence - Anti-GFP antibody [LGB-1] (ab291)This image is courtesy of an Abreview submitted by Vladimir Milenkovicab291 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.
-
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
文献 (24)
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