Anti-GFP tag Rabbit pAb - ChIP, CUT&RUN and CUT&Tag Grade

Product

Anti-GFP tag Rabbit pAb - ChIP, CUT&RUN and CUT&Tag Grade

Catalog number :AT0044

Green fluorescence protein (GFP) is a 27 kDa protein derived from jellyfish Aequorea victoria. It emits green light (emission peak at a wavelength of 509 nm) when excited by blue light (excitation peak at a wavelength of 395 nm). GFP has become a very useful tool as a fusion protein to report gene expression, trace cell lineage and define subcellular protein localizations. YFP differs from GFP due to a mutation at T203Y.

Overview

Description: Anti-GFP Rabbit pAb, ChIP, CUT&RUN and CUT&Tag Grade

Reactivity: Anti-GFP Antibody detects GFP-tagged proteins within cell lysates

Tested applications: WB (1:1000-5000), IP (1:100-300), IF (1:150-500), Flow Cyt, IHC-P, IHC-Fr

ChIP, CUT&RUN and CUT&Tag

Optimal dilutions/concentrations should be determined by the end user.

Product Picture

Properties

Immunogen: Highly purified recombinant full length protein made in Escherichia coli. The antibody is directed against the entire GFP molecule.

Clonality: Polyclonal (A mixture of 10 monoclonal antibodies )

This is to be more suitable for capturing the different conformational epitopes of GFP fusion protein exposed in chromatin for ChIP, CUT&RUN and CUT&Tag

Isotype: Rabbit IgG

Form: Liquid

Storage instruction: Store at -20°C, Avoid freeze / thaw cycle. Do not aliquot the antibody.

Applications

WB Image: Anti-GFP antibody (AT0044) at 1/2000 dilution + Lysate prepared from rabbit reticulocytes at 30 µg

Developed using the ECL technique

Performed under reducing conditions.

Observed band size: 27 kDa

Exposure time: 5 seconds

Recommended secondary antibody: Goat anti rabbit IgG-HRP (Engibody, Catalog number:AT0097)

Recommended ECL: ECL Pico-Detect™ Western Chemiluminescent HRP Substrate (Engibody, Catalog number: IF6747)

IHC Image: AT0044 staining dog hearts (Adv-GFP injection) tissue sections by IHC-P. Sections were PFA fixed and subjected to heat mediated antigen retrieval in citric acid (Ph6.0, 0.05% Tween20) prior to blocking with 10% serum for 30 mins at 37°C. The primary antibody was diluted 1/500 in PBS and incubated with the sample for 1 hour at 25°C.

ICC/IF Image: Immunofluorescence images showing similar localization of Yes-GFP (first 10 aa's of Yes PTK fused to the N-terminus of GFP) to full length Yes PTK.

A: Distribution of Yes detected using mouse anti-Yes Ab followed by Texas Red-conjugated anti-mouse pAb.

B: Chimeric GFP's detected using rabbit anti-GFP rabbit pAb (CAT: AT0044) followed by FITC-conjugated anti-rabbit pAb.

IP/CoIP Image: AT0044 Immunoprecipitate in human HEK 293 cells transfected with Annexin1-GFP. 25µg of cell lysate.

Line 1: Lysate of HEK 293 cells expressing Annexin1-GFP fusion

Line 2: IP with anti GFP pAb

Line 3: Not bound fraction

Anti-GFP tag Rabbit pAb - ChIP, CUT&RUN and CUT&Tag Grade

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