Citations et références (3)

Invitrogen™

InVision™ His-Tag In-Gel Staining Kit

Le kit de coloration InVision marquée à l’histidine sur gel comprend la coloration InVision marquée à l’histidine sur gel ainsiAfficher plus

Référence	Quantité
LC6033	1 kit

Référence LC6033

Prix (EUR)

1 260,00

Quantité:

1 kit

Grand volume ou format personnalisé

Prix (EUR)

1 260,00

Le kit de coloration InVision marquée à l’histidine sur gel comprend la coloration InVision marquée à l’histidine sur gel ainsi que l’étalon de protéines marquées à l’histidine BenchMark. La coloration InVision sur gel marquée à l’histidine est un colorant prêt à l’emploi, rapide, sensible et hautement spécifique pour visualiser les bandes de protéines de fusion marquées à l’histidine directement dans un gel de polyacrylamide après électrophorèse.

Caractéristiques :
• Colore les protéines fusionnées à la séquence d’oligo-histidine
• Élimine la détection par transfert Western : détection des protéines marquées à l’histidine directement dans le gel
• Détection des niveaux de nanogrammes des bandes de protéines avec un transilluminateur UV ou un équipement d’imagerie équipé de filtres adaptés ou de laser
• Visualisation des résultats en moins de trois heures

Comparez l’ensemble des colorations marquées à l’histidine ›

Une coloration rapide et facile
Le protocole de coloration marquée à l’histidine InVision sur gel est facile à exécuter et nécessite très peu de temps de manipulation, ce qui permet une sélection rapide et évolutive de l’expression des protéines dans une variété de types de gel. Après électrophorèse, le gel est fixé, incubé dans la coloration, lavé et la protéine de fusion spécifique marquée à l’histidine est détectée en exposant le gel aux UV ou à la lumière visible. En moins de trois heures, des niveaux de nanogramme de la protéine de fusion marquée à l’histidine peuvent être détectés. Après avoir documenté votre signal de coloration InVision, la coloration totale des protéines peut être effectuée.

Usage exclusivement réservé à la recherche. Ne pas utiliser pour des procédures de diagnostic.

Spécifications

Emplacement de détectionDétection sur gel

Méthode de détectionFluorescent

Quantité1 kit

Molécule cibleProtéines (marquées His)

Gamme de produitsInVision

Type de produitKit de coloration de protéines dans le gel sur marqueur His Kit

Unit SizeEach

Contenu et stockage

Invision™ His-tag In-gel est fournie sous la forme d’un réactif de coloration prêt à l’emploi 1X (500 ml).L’étalon protéique marqué à l’his BenchMark™ (125 μL) est également inclus dans le kit.

Foire aux questions (FAQ)

I used the InVision His-Tag In-Gel Stain to stain my His-Tagged fusion protein and used the BenchMark His-tagged protein standard as a positive control. I am seeing some additional bands with the BenchMark His-tagged Protein Standard. What went wrong?

This is likely due to overexposure - performing a longer exposure to detect low expression levels of the desired protein may result in staining of minor contaminants in the BenchMark His-tagged Protein Standard. Load less BenchMark His-tagged protein Standard or perform a short exposure to visualize and image the standard and then perform a longer exposure to visualize and image proteins expressed at low levels.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

I used the InVision His-Tag In-Gel Stain and am getting high, uneven background. Can you please offer some tips?

Here are possible causes and solutions:

- Missed washing steps. Be sure to wash the gel twice with 20 mM phosphate buffer. If the background is high, perform a third water wash step for 10 minutes.
- Poor water quality. Use ultrapure water (>18 megohm/cm) for washing and preparing phosphate buffer.
- Protein overloaded. Decrease the protein concentration or lower the sample volume.
- Dirty imaging platform. Always clean the imaging system with a paper towel prior to imaging the gel to minimize any background fluorescence.
- Non-specific bands. Highly basic proteins and divalent metal binding proteins such as carbonic anhydrase (30 kDa), SlyD (21 kDa), and phosphorylase B (97 kDa) may cross-react with the stain producing non-specific bands.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

I used the InVision His-Tag In-Gel Stain and am getting very weak staining. Can you please offer some tips?

Here are possible causes and solutions:

- Inadequate staining. Use appropriate staining protocol based on the gel type. Use BenchMark His-tagged Protein Standard as a positive control to verify staining reagents and protocol. Avoid excessive washing of the gel.
- The gel is not visualized or imaged properly. Be sure to visualize the gel using a UV transilluminator equipped with a camera or a laser-based scanner using the correct filters (see manual for details). A Polaroid camera is not recommended. Make sure the aperture on the camera is open wide to allow enough light entry and that the camera is connected to imaging software that allows contrast adjustment for viewing the best image. Visualize the gel immediately after completing the washing steps. Storing the gel in phosphate buffer decreases the signal intensity.
- Low protein load or expression level. Check total protein content of the gel by staining the gel with a total protein stain (check page 13 of the manual). Load at least 1 pmole of the His-tagged fusion protein for detection. Make sure the His-tag is in-frame and the protein is expressed properly.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Can I stain E-PAGE gels with InVision His-Tag In-Gel Stain?

E-PAGE gels are thicker than standard mini-gels and result in too much background when stained with InVision His-Tag In-Gel Stain. To obtain better staining sensitivity, we recommend transferring proteins of E-PAGE gels onto a nitrocellulose membrane and then staining the blot with the InVision His-tag In-gel Stain as described on page 14 in the manual.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

I have stained my His-tagged fusion protein with InVision His-Tag In-Gel Stain. Can I perform western blotting of the stained protein?

To perform western blotting after InVision His-Tag In-Gel staining of His-tagged fusion proteins:

- Record a permanent image of the gel after staining of His-tagged fusion proteins.
- Equilibrate the gel in 1X SDS Running Buffer for 1 hour.
- Perform western blotting and immunodetection using a method of choice.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

View all InVision™ His-Tag In-Gel Staining Kit FAQs

Citations et références (3)

Citations et références

Abstract

Novel determinant of PKC-epsilon anchoring at cardiac Z-lines.

Authors:Robia SL, Kang M, Walker JW,

Journal:Am J Physiol Heart Circ Physiol

PubMed ID:15964917

'The Z-line represents a critical link between the transverse tubule network and cytoskeleton of cardiac cells with a role in anchoring structural proteins, ion channels, and signaling molecules. Protein kinase C-epsilon (PKC-epsilon) regulates cardiac excitability, cardioprotection, and growth, possibly as a consequence of translocation to the Z-line/T tubule region. To ... More

Dss1 interaction with Brh2 as a regulatory mechanism for recombinational repair.

Authors:Zhou Q, Kojic M, Cao Z, Lisby M, Mazloum NA, Holloman WK,

Journal:Mol Cell Biol

PubMed ID:17261595

Brh2, the BRCA2 ortholog in Ustilago maydis, enables recombinational repair of DNA by controlling Rad51 and is in turn regulated by Dss1. Interplay with Rad51 is conducted via the BRC element located in the N-terminal region of the protein and through an unrelated domain, CRE, at the C terminus. Mutation ... More

A conserved inverted repeat, the LipR box, mediates transcriptional activation of the Streptomyces exfoliatus lipase gene by LipR, a member of the STAND class of P-loop nucleoside triphosphatases.

Authors:Evangelista-Martínez Z, González-Cerón G, Servín-González L,

Journal:J Bacteriol

PubMed ID:17015647

Expression of the Streptomyces exfoliatus lipA gene, which encodes an extracellular lipase, depends on LipR, a transcriptional activator that belongs to the STAND class of P-loop nucleoside triphosphatases. LipR is closely related to activators present in some antibiotic biosynthesis clusters of actinomycetes, forming the LipR/TchG family of regulators. In this ... More