c-MYC anticorps (AA 31-80)
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- Antigène Voir toutes c-MYC (MYC) Anticorps
- c-MYC (MYC) (Myc Proto-Oncogene protein (MYC))
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Épitope
- AA 31-80
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Reactivité
- Humain, Souris, Rat
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Hôte
- Lapin
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Clonalité
- Polyclonal
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Conjugué
- Cet anticorp c-MYC est non-conjugé
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Application
- Western Blotting (WB), Immunohistochemistry (IHC), ELISA, Immunofluorescence (IF)
- Specificité
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MYC Antibody detects endogenous levels of total MYC protein.
- Purification
- The antibody was purified from rabbit antiserum by affinity-chromatography using immunogen.
- Pureté
- > 95 %
- Immunogène
- The antiserum was produced against synthesized peptide derived from human MYC.
- Isotype
- IgG
- Top Product
- Discover our top product MYC Anticorps primaire
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- Indications d'application
- WB: 1:500~1:1000 IHC: 1:50~1:100 IF: 1:100~1:500 ELISA: 1:40000
- Commentaires
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Unigene-Number: Hs.202453 (NCBI Gene Symbol: MYC)
- Restrictions
- For Research Use only
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- by
- Developmental Biology, Johann-Friedrich-Blumenbach-Institute for Zoology and Anthropology, Georg-August-University of Göttingen
- No.
- #102753
- Date
- 03.07.2018
- Antigène
- MYC
- Numéro du lot
- 210020 and 424170020
- Application validée
- Western Blotting
- Contrôle positif
HEK-293 cells transfected with Myc expression plasmid
- Contrôle négative
HEK-293 cells transfected with HA-Spag4-myc expression plasmid
- Conclusion
Passed. ABIN1532205 detects Myc protein in a western blot. No cross-reaction was observed with the myc-tag sequence positioned at the C-terminus of a recombinant protein.
- Anticorps primaire
- ABIN1532205
- Anticorps secondaire
- rabbit anti-mouse IgG, HRP-linked (Sigma A9044, Lot 034M4761)
- Full Protocol
- Grow HEK-293 cells (ATCC, CRL-1658) in DMEM+GlutaMAXDMEM+GlutaMAX (Gibco, 10567-014, lot 1922818) supplemented with fetal bovine serum (Gibco, 10500-064, lot 08FO477K) and Pen/Strep (Gibco, 15140-122, lot 1924797) at 37°C and 5% CO2 to 70% confluency.
- Transfect cells either with a Myc expression plasmid (pcDNA3.1-Myc) or HA- and myc-tagged SPAG4 expression plasmids using EndofectinMax (GeneCopoeia) following the manufacturer´s instructions.
- Grow cells for 24h.
- Lyse cells in SDS-sample buffer and denature total cellular lysates for 5min at 95°C. Subsequently separate them on a denaturing 10% SDS-PAGE (Laemmli 1970).
- Transfer proteins onto 0.2µm Protran membrane (GE Healthcare, 10600004, A10043108) by wet blotting for 1h at 400mA (Towbin et al., 1979).
- Block the membrane in TBST (50mM Tris-HCl, pH7.4, 150mM NaCl, 0.2% Tween 20) containing 5% milk (blocking solution) for 60min at RT.
- Incubate membrane with primary rabbit anti-myc antibody (antibodies-online, ABIN1532205 Lot#424170020 and Lot#210020) diluted 1:500 in blocking solution overnight at 4°C.
- Wash membrane with TBST for 45min at RT.
- Incubate membrane with secondary goat anti-rabbit IgG (H+L), HRP-linked (Jackson Immuno Research, 111-035-003, lot 123450) diluted 1:1000 in blocking solution for 45min at RT.
- Wash membrane 6x for 5min with TBST.
- Reveal protein bands using Clarity Max Western ECL substrate (Bio-Rad, 1705062) and capture images via Chemidoc Imaging System (BioRad).
- Incubate membrane with primary mouse anti-HA-tag antibody (clone 12CA5) diluted 1:500 in blocking solution overnight at 4°C.
- Wash membrane with TBST for 45min at RT.
- Incubate membrane with secondary rabbit anti-mouse IgG, HRP-linked (Sigma A9044, Lot 034M4761) diluted 1:1000 in blocking solution for 45min at RT.
- Reveal protein bands using Clarity Max Western ECL substrate (Bio-Rad, 1705062).
- Notes
In lysates of HEK-293 cells ectopically expressing recombinant protein with an N-terminal HA- and a C-terminal myc-tag ABIN1532205 did not detect the labeled protein at the expected molecular weight. In contrast, an anti-HA-tag antibody (clone 12CA5) revealed a protein at the expected MW.
Validation #102753 (Western Blotting)Validation ImagesProtocole -
- by
- Developmental Biology, Johann-Friedrich-Blumenbach-Institute for Zoology and Anthropology, Georg-August-University of Göttingen
- No.
- #103023
- Date
- 03.07.2018
- Antigène
- MYC
- Numéro du lot
- 210020 and 424170020
- Application validée
- Immunocytochemistry
- Contrôle positif
HEK-293 cells transfected with Myc expression plasmid
- Contrôle négative
HEK-293 cells transfected with HA-Spag4-myc expression plasmid
- Conclusion
Passed. ABIN1532205 (lot 424170020 and lot 210020) detects Myc in immunocytochemistry. No cross-reaction was observed with the myc-tag sequence positioned at the C-terminus of a recombinant protein.
- Anticorps primaire
- ABIN1532205
- Anticorps secondaire
- goat anti-rabbit IgG MFP590 (Mobitec MFP-A1037, lot 3002088), F(ab')2-Goat anti-Rabbit IgG (H+L) Alexa Fluor 555 (Life Technologies, A21430, lot 1637266)
- Full Protocol
- Grow HEK-293 cells (ATCC, CRL-1658) in DMEM+GlutaMAXDMEM+GlutaMAX (Gibco, 10567-014, lot 1922818) supplemented with fetal bovine serum (Gibco, 10500-064, lot 08FO477K) and Pen/Strep (Gibco, 15140-122, lot 1924797) at 37°C and 5% CO2 to 70% confluency.
- Transfect cells either with a Myc expression plasmid (pcDNA3.1-Myc) or HA- and myc-tagged SPAG4 expression plasmids using EndofectinMax (GeneCopoeia) following the manufacturer´s instructions.
- Grow cells for 24h.
- Fix cells in 3.7% paraformaldehyde (in PBS) for 20min at 4°C followed by incubation in 0.3% Triton X-100 for 10min at 4°C.
- Block cells in PBS containing 1% BSA and 0.5% Tween-20 (PBT) for 1h at RT.
- Incubate cells with primary antibody
- rabbit anti-Myc antibody (antibodies-online, ABIN1532205, lot 424170020 or lot 210020) diluted 1:50 in PBS and mouse anti-HA tag antibody (clone 12CA5) ON at 4°C (images G, H).
- rabbit anti-Myc antibody (antibodies-online, ABIN1532205, lot 210020 or lot 210020) diluted 1:50 in PBS ON at 4°C (images A, D).
- Wash cells with 50mM Tris-HCl, pH7.4, 150mM NaCl, 0.1% Tween 20 (TBST) for 15min at RT.
- Incubate cells with secondary antibody
- goat anti-rabbit IgG MFP590 (Mobitec MFP-A1037, lot 3002088) diluted 1:300 in PBS and goat anti-mouse IgGDylight488 (Thermo Scientific, 35503) diluted 1:10000 in PBS for 1h at 37°C (images G, H).
- F(ab')2-Goat anti-Rabbit IgG (H+L) Alexa Fluor 555 (Life Technologies, A21430, lot 1637266) diluted 1:2000 in PBS for 45min at 37°C (images A, D).
- Counterstain cells with DAPI (Sigma, D-9542).
- Image acquisition on Zeiss LSM 510 confocal microscope and processing using Adobe Photoshop 5.0.
- Notes
The antibody detects Myc proteins by immunoblotting and by immunocytology. No cross-reaction with the C-terminal Myc-tag.
Validation #103023 (Immunocytochemistry)Validation ImagesABIN1532205 detects ectopically expressed Myc (pcDNA3.1-Myc) (A, red). No cross-reactivity is observed in untransfected cells (D). DAPI was used to visualize the nucleus (B and E, blue). Images on the right show the merged red and blue channels. ABIN1532205 did not detect the myc-tag (G, red) of HA-Spag4-myc whereas an anti HA-tag antibody (H, green) detected HA-Spag4-myc at the nuclear membrane as expected. The picture on the right (J) shows the red and green channels merged with DAPI counterstain (I, blue).Protocole -
- Format
- Liquid
- Concentration
- 1 mg/mL
- Buffer
- phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150 mM NaCl, 0.02 % sodium azide and 50 % glycerol.
- 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.
- Stock
- -20 °C
- Stockage commentaire
- Stable at -20°C for at least 1 year.
- Date de péremption
- 12 months
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- Antigène
- c-MYC (MYC) (Myc Proto-Oncogene protein (MYC))
- Autre désignation
- Myc Proto-Oncogene protein (MYC Produits)
- Synonymes
- anticorps MRTL, anticorps MYCC, anticorps bHLHe39, anticorps c-Myc, anticorps CMYC, anticorps c-myc, anticorps C-MYC, anticorps LOC100136746, anticorps ARC, anticorps B430311C09Rik, anticorps MYC, anticorps NOP, anticorps Nop30, anticorps RNCMYC, anticorps mMyc, anticorps AU016757, anticorps Myc2, anticorps Niard, anticorps Nird, anticorps cmyc, anticorps zc-myc, anticorps c-myc II, anticorps myc-B, anticorps myc2, anticorps myc1, anticorps CMyc, anticorps zgc:55680, anticorps CG10798, anticorps D-Myc, anticorps DM, anticorps DMYc, anticorps Dm, anticorps Dmel\CG10798, anticorps Dmyc, anticorps EG:BACN5I9.1, anticorps anon-WO03040301.171, anticorps bHLHe57, anticorps c-MYC, anticorps d-myc, anticorps dMYC, anticorps dMyc, anticorps dMyc1, anticorps da, anticorps dm, anticorps dm/dMyc, anticorps dm/myc, anticorps dmyc, anticorps dmyc1, anticorps l(1)G0139, anticorps l(1)G0354, anticorps l(1)G0359, anticorps myc, anticorps MYC proto-oncogene, bHLH transcription factor, anticorps v-myc myelocytomatosis viral oncogene homolog, anticorps nucleolar protein 3 (apoptosis repressor with CARD domain), anticorps myelocytomatosis oncogene, anticorps MYC proto-oncogene, bHLH transcription factor a, anticorps MYC proto-oncogene, bHLH transcription factor L homeolog, anticorps MYC proto-oncogene, bHLH transcription factor S homeolog, anticorps v-myc myelocytomatosis viral oncogene homolog (avian), anticorps MYC proto-oncogene, bHLH transcription factor b, anticorps CG10798 gene product from transcript CG10798-RB, anticorps MYC, anticorps LOC100136746, anticorps Nol3, anticorps Myc, anticorps myca, anticorps myc.L, anticorps myc.S, anticorps mycb
- Sujet
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Synonyms: Myc proto-oncogene protein, Class E basic helix-loop-helix protein 39, bHLHe39, Proto-oncogene c-Myc, Transcription factor p64 , MYC , BHLHE39
NCBI Gene Symbol: MYC
- Poids moléculaire
- 48 kDa
- ID gène
- 4609
- OMIM
- 113970
- UniProt
- P01106
- Pathways
- Signalisation p53, Cycle Cellulaire, Sensory Perception of Sound, Transition Metal Ion Homeostasis, Mitotic G1-G1/S Phases, Positive Regulation of Endopeptidase Activity, Regulation of Carbohydrate Metabolic Process, Positive Regulation of Response to DNA Damage Stimulus, L'effet Warburg
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