Product datasheet

Anti-GFP antibody ab13970
93 Abreviews 3610 References 画像数 8
医薬用外劇物

製品の概要
製品名

Anti-GFP antibody

製品の詳細

Chicken polyclonal to GFP

由来種Chicken

特異性

Our GFP antibody does cross-react with the many fluorescent proteins that are derived from thejellyfish Aequorea victoria. These are all proteins that differ from the original GFP by just a few point
mutations (EGFP, YFP, mVenus, CFP, BFP etc.).

アプリケーション

適用あり : WB, ICC/IF

種交差性

交差種 : Species independent

免疫原Recombinant full length protein corresponding to GFP.
Database link: P42212

ポジティブ･コントロール

ICC: GFP-transfected NIH/3T3 (Mouse embryo fibroblast cell line). WB: Transgenic mouse spinalcords.

特記事項

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 befound below, along with publications, customer reviews and Q&As

製品の特性
製品の状態

Liquid

保存方法

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C.
Avoid freeze / thaw cycle.

バッファー

pH: 7
Preservative: 0.01% Thimerosal (merthiolate)
Constituents: PBS, 50% Glycerol, 0.16% Sodium phosphate

精製度

IgY fraction

特記事項（精製）

Sterile filtered.
1ポリ /モノ

ポリクローナルアイソタイプIgY

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab13970の使用に適用されますアプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション特記事項

Abreviews

WB

(13)

1/5000.

ICC/IF(22)

1/2000.
Used at a dilution of 1/2000 for 1 hr (see Abreview for furtherinformation).

ターゲット情報
関連性

Function: Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of theprotein 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 acidresidues. 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- andC-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, reporterof gene expression, or as a measure of protein-protein interactions. Can also be used as amolecular 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.

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画像

All lanes : Anti-GFP antibody (ab13970) at 1/2000 dilution (Diluent
1x TBS /4 hours at 4°C)

Lane 1 : 3 µg of GFP plasmid overexpressed in mousecardiomyocytes whole cell lysate with BSA / for 1 hour at room
temperature

Lane 2 : 2 µg of GFP plasmid overexpressed in mousecardiomyocytes whole cell lysate with BSA / for 1 hour at room
temperature

Lane 3 : 1 µg of GFP plasmid overexpressed in mousecardiomyocytes whole cell lysate with BSA / for 1 hour at room
Western blot - Anti-GFP antibody (ab13970)

temperatureThis image is courtesy of an anonymous Abreview

Lysates/proteins at 25 µg per lane.
Blocking peptides at 5 % per lane.

SecondaryAll lanes : Goat Anti-Chicken IgY H&L (Alexa Fluor® 594)
preadsorbed (ab150176) at 1/5000 dilution
Performed under reducing conditions.

Additional bands at: 25 kDa. We are unsure as to the identity ofthese extra bands.

Exposure time: 30 seconds

Gel Running Conditions: Reduced Denaturing (15% PAGE)
Detection method: Fluorescent Secondary Antibodies3ab13970 staining GFP in U-2 OS (Human bone osteosarcoma
epithelial cell line) cells by ICC/IF.
Cells were paraformaldehyde fixed, permeabilized with 0.5%
triton and blocked with 2% antibody dilution buffer for 2 hours. Cells
were incubated with the primary antibody (1/1000) for 1 hour at 25°C.
An undiluted Alexa Fluor® 488 conjugated Goat anti-chicken
polyclonal was used as the secondary antibody.

Immunocytochemistry/ Immunofluorescence - AntiGFP antibody (ab13970)
This image is courtesy of an Abreview submitted byChristophe Lachaud

All lanes : Anti-GFP antibody (ab13970) at 1/1000 dilution
Lane 1 : Whole cell lysate prepared from HeLa (Human epithelial
cell line from cervix adenocarcinoma) cells.

Lane 2 : Whole cell lysate prepared from HeLa cells.
Lysates/proteins at 25 µg per lane.

SecondaryAll lanes : IRDye 800CW conjugated goat anti-chicken polyclonal
at 1/15000 dilution
Western blot - Anti-GFP antibody (ab13970)
Image courtesy of an anonymous Abreview.

Transgenic mice expressing GFP selectively in lamina II of the spinalcord.
In the right panels, note the correspondance between the green(rabbit anti-GFP) and red signals (chicken anti-GFP from Abcam)
indicating that these two antibody preparations recognized the samegene product. The secondary antibody used with ab13970 was an
FITC-labeled goat anti-chicken

Immunocytochemistry/ Immunofluorescence - AntiGFP antibody (ab13970)

4ab13970 staining GFP in GFP-transfected NIH/3T3 (Mouse embryo
fibroblast cell line) cells.
The cells were fixed with 4% formaldehyde (10 minutes) and thenblocked in 1% BSA / 0.3M glycine in 0.1%PBS-Tween for 1 hour.
The cells were then incubated with ab13970 at 1/2000 dilution
overnight at +4°C followed by incubation with Goat Anti-Chicken IgY
H&L (Alexa Fluor® 488) preadsorbed (ab150173), for 1 hour, at 1μg/ml.
Under identical experimental conditions, when compared to the basal
level of GFP expression in transfected NIH/3T3 cells, the cells uponwhich ab13970 was applied gave a stronger signal in the 488
Immunocytochemistry/ Immunofluorescence - Anti-channel, indicating that ab13970 is binding to GFP and therefore

GFP antibody (ab13970)

eliciting signal amplification.
ab13970 was also applied to non-GFP-transfected NIH/3T3 cells,
which produced no positive staining, indicating specificity for GFP.
Nuclear DNA was labeled with 1.43 μM DAPI (blue).

Lane 1 : Rabbit anti-GFP
Lane 2 : Anti-GFP antibody (ab13970)
All lanes : Transgenic mouse spinal cords

Western blot - Anti-GFP antibody (ab13970)

Western blot of transgenic mouse spinal cords showing that therabbit anti-GFP (lane 1) and the chicken anti-GFP (Abcam; lane 2)
recognize a band at the same molecular weight.

ab13970 staining GFP + tumor in mouse muscle cells by ICC/IF.
Cells were formaldehyde fixed and blocked with 3% BSA for 1 hour
at 24°C prior to incubation with the primary antibody (1/500) for 1
hour at 24°C. An Alexa Fluor® 488 conjugated goat anti-chicken wasused as the secondary.

Immunocytochemistry/ Immunofluorescence - AntiGFP antibody (ab13970)
This image is courtesy of an Abreview submitted by Dr
Radbod Darabi

5Immunocytochemical/ Immunoflurescence analysis of cytospined
HEK-293 cells (Human epithelial cell line from embryonic kidney)transfected with GFP, labeling GFP with ab13970 at 1/200 incubatedfor 16 hours at 4°C with 1% BSA in PBS. Secondary used was adonkey anti-chicken polyclonal DyLight® 594 at 1/500.
GFP is shown in red (DyLight® 594).
Nuclei are counterstained in blue (DAPI).
The left panel shows HEK-293 cells transfected with GFP and theImmunocytochemistry/ Immunofluorescence - Anti-right panel shows non-transfected HEK-293 cells.
GFP antibody (ab13970)
Image courtesy of an AbReview submitted by Dr
Francois Daubeuf.

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