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PA1-980A - Western blot analysis of recombinant GFP was performed by loading 10 uL (250 ng) of samples and 7ul of Molecular Weight Protein Ladder per well onto a 4-20% Bis-Tris polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and stained/destained with Pierce MemCode Blue Kit (Product #24580). Membranes were probed with Pierce Fast Western Kit (Product #35050). Proteins were stained with rabbit polyclonal antibodies recognizing GFP (Product #PA1980A). PA1980A, lot MF153968 at 0.5 ug/mL (Lanes 8); PA1980A, lot NL174160 at 1.0 ug/mL (Lane 9), 0.5 ug/mL (Lane 10) and 0.25 ug/mL (Lane 11); and PA1980A, lot NL174160 at 0.5 ug/mL with 2.5 ug/mL blocking, immunizing peptide (Lane 12) were incubated for 30 minutes at room temperature on a rocking platform per Fast Western Kit instructions. Membranes were then washed in kit wash buffer and probed with kit secondary antibody reagent for 10 minutes diluted at 150 uL per 10 mL kit reagent diluent. Membranes were washed in kit wash buffer and chemiluminescent detection was performed using Pierce Super Signal West Dura (Product #34075).
|Tested species reactivity||Tag|
|Published species reactivity||Rat, Pig, Plant, Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: H(26) K F S V S G E G E G D A T(39) C|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||0.5 ug/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-980A detects Green Fluorescent Protein (GFP).
PA1-980A has been successfully used in Western blot and immunofluoresence procedures. By Western blot, this antibody detects recombinant GFP.
PA1-980A immunizing peptide corresponds to the N-terminal amino acid residues 26-39 of Aequorea victoria wild-type GFP. This sequence is completely conserved between GFP and the modified and enhanced Blue, Green, and Yellow FP variants. PA1-980A immunizing peptide (Cat. # PEP-033) is available for use in neutralization and control experiments.
Green Fluorescent Protein (GFP) is responsible for bioluminescence in the jellyfish Aequorea victoria. This protein has quickly become a powerful research tool for assessing gene expression and subcellular protein distribution in fixed or living cells. GFP is excited by and brightly fluoresces when exposed to UV or blue light. This feature makes it ideal as a marker for use in fluorescence microscopy, cytometry, tagging fusion proteins, and assaying transcriptional regulation from gene promoters in vivo. Numerous GFP variants with enhanced and shifted emission spectra (blue, green, and yellow) have been developed through amino acid substitutions at specific residues.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
|Not Applicable||Not Cited||
Probing protein interactions in living mammalian cells on a microtubule bench.
PA1-980A was used in western blot to develop a method to use microtubules as platforms to detect protein interactions in mammalian cells
|Boca M,Kretov DA,Desforges B,Mephon-Gaspard A,Curmi PA,Pastré D||Scientific reports (5:null)||2015|
|Not Applicable||Not Cited||
Liver X receptor ligand cytotoxicity in colon cancer cells and not in normal colon epithelial cells depends on LXRß subcellular localization.
PA1-980A was used in western blot to investigate LXRbeta signaling in colon cancer cells
|Courtaut F,Derangère V,Chevriaux A,Ladoire S,Cotte AK,Arnould L,Boidot R,Rialland M,Ghiringhelli F,Rébé C||Oncotarget (6:26651)||2015|
Heterodimers of tyrosylprotein sulfotransferases suggest existence of a higher organization level of transferases in the membrane of the trans-Golgi apparatus.
PA1-980A was used in immunocytochemistry to investigate the localization and binding partners of the tyrosyl protein sulfotransferases, TPST1 and TPST2
|Hartmann-Fatu C,Trusch F,Moll CN,Michin I,Hassinen A,Kellokumpu S,Bayer P||Journal of molecular biology (427:1404)||2015|
Development of a tightly regulated and highly responsive copper-inducible gene expression system and its application to control of flowering time.
PA1-980A was used in western blot to study the ability to modulate flowering time using a novel copper-inducible expression system
|Saijo T,Nagasawa A||Plant cell reports (33:47)||2014|
Unfolding-resistant translocase targeting: a novel mechanism for outer mitochondrial membrane localization exemplified by the Bbeta2 regulatory subunit of protein phosphatase 2A.
PA1-980A was used in western blot to study the localization of heterotrimeric serine/threonine protein phosphatase 2A beta2 subunit.
|Dagda RK,Barwacz CA,Cribbs JT,Strack S||The Journal of biological chemistry (280:27375)||2005|
An amino-terminal motif functions as a second nuclear export sequence in BRCA1.
PA1-980A was used in western blot to identify nuclear export sequences in BRAC1 gene .
|Thompson ME,Robinson-Benion CL,Holt JT||The Journal of biological chemistry (280:21854)||2005|
A novel method for the production of transgenic cloned pigs: electroporation-mediated gene transfer to non-cultured cells and subsequent selection with puromycin.
PA1-980A was used in western blot to evaluate a novel way to produe transgenic cloned pigs
|Watanabe S,Iwamoto M,Suzuki S,Fuchimoto D,Honma D,Nagai T,Hashimoto M,Yazaki S,Sato M,Onishi A||Biology of reproduction (72:309)||2005|
|Not Applicable||10 ug/ml||
Development of a continuous assay for the measurement of tissue factor procoagulant activity on intact cells.
PA1-980A was used in blocking or activating experiment to evaluate a novel assay for the continuous measurement of tissue factor procoagulant activity in intact cells
|Caldwell JA,Dickhout JG,Al-Hashimi AA,Austin RC||Laboratory investigation; a journal of technical methods and pathology (90:953)||2010|
eGFP; GFP; GFP2; Green Fluoresecent Protein; Turbo GFP