GRK6 anticorps (C-Term)
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- Antigène Voir toutes GRK6 Anticorps
- GRK6 (G Protein-Coupled Receptor Kinase 6 (GRK6))
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Épitope
- AA 382-417, C-Term
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Reactivité
- Humain, Rat
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Hôte
- Lapin
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Clonalité
- Polyclonal
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Conjugué
- Cet anticorp GRK6 est non-conjugé
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Application
- Western Blotting (WB)
- Fonction
- Rabbit IgG polyclonal antibody for G protein-coupled receptor kinase 6(GRK6) detection. Tested with WB in Human,Rat.
- Séquence
- QSPFQQRKKK IKREEVERLV KEVPEEYSER FSPQAR
- Réactivité croisée (Details)
- No cross reactivity with other proteins.
- Attributs du produit
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Rabbit IgG polyclonal antibody for G protein-coupled receptor kinase 6(GRK6) detection. Tested with WB in Human,Rat.
Gene Name: G protein-coupled receptor kinase 6
Protein Name: G protein-coupled receptor kinase 6 - Purification
- Immunogen affinity purified.
- Immunogène
- A synthetic peptide corresponding to a sequence at the C-terminus of human GRK6 (382-417aa QSPFQQRKKKIKREEVERLVKEVPEEYSERFSPQAR), different from the related mouse sequence by four amino acids, and from the related rat sequence by three amino acids.
- Isotype
- IgG
- Top Product
- Discover our top product GRK6 Anticorps primaire
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- Indications d'application
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WB: Concentration: 0.1-0.5 μg/mL, Tested Species: Human, Rat
Notes: Tested Species: Species with positive results.
Other applications have not been tested. Optimal dilutions should be determined by end users. - Commentaires
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Antibody can be supported by chemiluminescence kit ABIN921124 in WB.
- Restrictions
- For Research Use only
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- by
- Institut für Molekulare Zellbiologie, Universitätsklinikum Jena
- No.
- #103590
- Date
- 31.01.2019
- Antigène
- GRK6
- Numéro du lot
- 0971512Da9609109
- Application validée
- Western Blotting
- Contrôle positif
HEK293 cells transiently transfected with pcDNA3 vectors containing GRK6 transcript variant 1 (NM_001004106.3), transcript variant 2 (NM_002082.3), transcript variant 3 (NM_001004105.2), or transcript variant 4 (NM_001364164.1)
- Contrôle négative
HEK293 cells transiently transfected with empty pcDNA3 vector
- Conclusion
The GRK6 antibody ABIN3042435 is able to detect all four tested GRK6 isoforms, but gives strong background-signals around 55kDa.
- Anticorps primaire
- ABIN3042435
- Anticorps secondaire
- goat-anti-rabbit Peroxidase-labeled antibody (SeraCare, 5220-0336)
- Full Protocol
- Grow 7.5x105 HEK293 cells in DMEM-medium (Sigma-Aldrich, D6429)) supplemented with 10% Fetal Bovine serum (Sigma-Aldrich, F7524) and 1% penicillin-streptomycin (Sigma-Aldrich, P0781), at 37°C and 5% CO2 in 2ml on a 6-well-dish (Greiner Bio-One) ON.
- Transfect cells with 2μg of respective pcDNA3 construct using self-prepared PEI transfection reagent.
- After 24h lyse the cells in 250μl per well cold RIPA lysis buffer (1 % NP-40, 1mM EDTA, 50mM Tris-HCl pH7.4, 150mM NaCl, 0.25 % Sodium-deoxycholate, PhosSTOP tablet (Roche, 04906845001) and cOmplete tablet (Roche, 04693132001) diluted in RIPA following the manufacturer´s instructions.
- Denature the cleared lysate of total protein for 5min at 95°C in 50μl of 6x SDS sample buffer and subsequently separate 4μl of each sample on a 10% polyacrylamide gel.
- Transfer proteins onto nitrocellulose membrane (Biostep 01-14-101) with a Tank Blotting System at 10V ON.
- Block the membrane with 1x Casein Blocking Buffer (Sigma-Aldrich, B6429) for 1h at RT with gentle shaking.
- Wash membrane 3x for 10min with TBST.
- Cut the membrane and incubate fragments separately with primary
- rabbit anti-GRK6 antibody (ABIN3042435, antibodies-online, lot 0971512Da9609109) diluted 1:1000 in 5% BSA-TBST at 4°C ON.
- rabbit anti-GRK6 antibody (Cell Signaling Technology, 5878, lot 1) diluted 1:1000 in 5% BSA-TBST at 4°C ON.
- rabbit anti-Vinculin antibody (Biozol, BZL03106, lot 0401) diluted 1:1000 in 5% BSA-TBST at 4°C ON.
- Wash membrane 3x for 10min with TBST.
- Incubate membrane with secondary goat-anti-rabbit Peroxidase-labeled antibody (SeraCare, 5220-0336) diluted 1:10000 in 1x Casein Blocking Buffer) for 1h at RT with gentle shaking.
- Wash membrane 3x for 10min with TBST.
- Reveal protein bands using Western Lightning Plus ECL reagent (Perkin Elmer, NEL103001EA, Luminol reagent Lot 275-17431, Oxidizing reagent, lot 265-17431) on a LAS-4000 Luminescence Imager (Fujifilm), exposure for 15 and 90 seconds.
- Notes
If the detection of the smallest isoform (GRK6-4, transcript variant 4, NM_001364164.1) is required, ABIN3042435 is a good choice, as this isoform is not detected by the alternative GRK6 antibody tested in parallel. Nevertheless, the strong background signal around 55kDa produced by ABIN3042435 diminishes its usefulness.
Validation #103590 (Western Blotting)Validation ImagesWestern blot analysis of HEK293 cell lysates subsequently to transient transfection with pcDNA3 constructs containing no insert (EV), GRK6 transcript variant 1, NM_001004106.3, transcript variant 2, NM_002082.3, transcript variant 3, NM_001004105.2, or transcript variant 4, NM_001364164.1 as indicated. Equal amounts of the lysates were loaded twice on the same gel and after blotting, the membrane was cut and incubated with ABIN3042435, a reference anti-GRK6 antibody (GRK6), and Vinculin loading control antibody (Vinculin) in parallel. Exposure for the upper panel was 15 seconds and for the lower 90 seconds.
Protocole -
- Format
- Lyophilized
- Reconstitution
- Add 0.2 mL of distilled water will yield a concentration of 500 μg/mL.
- Concentration
- 500 μg/mL
- Buffer
- Each vial contains 5 mg BSA, 0.9 mg NaCl, 0.2 mg Na2HPO4, 0.05 mg Sodium azide.
- Agent conservateur
- Sodium azide
- Précaution d'utilisation
- This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
- Conseil sur la manipulation
- Avoid repeated freezing and thawing.
- Stock
- 4 °C/-20 °C
- Stockage commentaire
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At -20°C for one year. After reconstitution, at 4°C for one month.
It can also be aliquotted and stored frozen at -20 °C for a longer time. Avoid repeated freezing and thawing.
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Suitability of GRK Antibodies for Individual Detection and Quantification of GRK Isoforms in Western Blots." dans: International journal of molecular sciences, Vol. 23, Issue 3, (2022) (PubMed).
: "
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Suitability of GRK Antibodies for Individual Detection and Quantification of GRK Isoforms in Western Blots." dans: International journal of molecular sciences, Vol. 23, Issue 3, (2022) (PubMed).
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- Antigène
- GRK6 (G Protein-Coupled Receptor Kinase 6 (GRK6))
- Autre désignation
- GRK6 (GRK6 Produits)
- Synonymes
- anticorps MGC83187, anticorps GRK6, anticorps GPRK6, anticorps Gprk6, anticorps G protein-coupled receptor kinase 6, anticorps G protein-coupled receptor kinase 6 S homeolog, anticorps GRK6, anticorps grk6.S, anticorps grk6, anticorps Grk6
- Sujet
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G protein-coupled receptor kinase 6 is an enzyme that in humans is encoded by the GRK6 gene. It is mapped to 5q35. This gene encodes a member of the guanine nucleotide-binding protein (G protein)-coupled receptor kinase subfamily of the Ser/Thr protein kinase family. The protein phosphorylates the activated forms of G protein-coupled receptors thus initiating their deactivation. Several transcript variants encoding different isoforms have been described for this gene. Also, GRK6 appears to be involved in responses to morphine.
Synonyms: FLJ32135 antibody|G protein coupled receptor kinase 6 antibody|G protein coupled receptor kinase GRK6 antibody|G protein-coupled receptor kinase 6 antibody|G protein-coupled receptor kinase GRK6 antibody|Gprk6 antibody|Grk6 antibody|GRK6_HUMAN antibody - ID gène
- 2870
- UniProt
- P43250
- Pathways
- Myometrial Relaxation and Contraction, Regulation of G-Protein Coupled Receptor Protein Signaling, CXCR4-mediated Signaling Events, Negative Regulation of Transporter Activity
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