MYC (AA 31-80) antibody

Details for Product No. ABIN1532205, Supplier: Log in to see
Antigen
  • RNCMYC
  • c-myc
  • mMyc
  • AU016757
  • Myc2
  • Niard
  • Nird
  • bHLHe39
  • MYC proto-oncogene, bHLH transcription factor
  • myelocytomatosis oncogene
  • Myc
Epitope
AA 31-80
38
34
24
17
17
16
10
10
9
5
4
3
3
3
3
3
3
3
3
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
Reactivity
Human, Mouse (Murine), Rat (Rattus)
317
109
102
7
3
3
3
3
3
2
1
1
1
1
1
1
Host
Rabbit
235
76
4
2
2
1
Clonality
Polyclonal
Conjugate
Un-conjugated
7
7
7
7
6
5
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
1
Application
Immunofluorescence (IF), Immunohistochemistry (IHC), ELISA, Western Blotting (WB)
243
136
107
86
45
34
30
28
17
14
10
4
3
1
1
Options
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'Independent Validation' Badge
Antigen MYC
Lot Number 210020 and 424170020
Method validated Western Blotting
Positive Control

HEK-293 cells transfected with Myc expression plasmid

Negative Control

HEK-293 cells transfected with HA-Spag4-myc expression plasmid

Notes

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.

Primary Antibody ABIN1532205
Secondary Antibody rabbit anti-mouse IgG, HRP-linked (Sigma A9044, Lot 034M4761)
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).
Experimental 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 Images
Western Blotting validation image for anti-MYC (AA 31-80) antibody (ABIN1532205) ABIN1532205 lot 424170020 (let panel) and lot 210020 (middle and right panel) detecte...
'Independent Validation' Badge
Antigen MYC
Lot Number 210020 and 424170020
Method validated Immunocytochemistry
Positive Control

HEK-293 cells transfected with Myc expression plasmid

Negative Control

HEK-293 cells transfected with HA-Spag4-myc expression plasmid

Notes

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.

Primary Antibody ABIN1532205
Secondary Antibody 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)
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.
Experimental Notes

The antibody detects Myc proteins by immunoblotting and by immunocytology. No cross-reaction with the C-terminal Myc-tag.

Validation Images
Immunocytochemistry validation image for anti-MYC (AA 31-80) antibody (ABIN1532205) ABIN1532205 detects ectopically expressed Myc (pcDNA3.1-Myc) (A, red). No cross-react...
Immunogen The antiserum was produced against synthesized peptide derived from human MYC.
Isotype IgG
Specificity

MYC Antibody detects endogenous levels of total MYC protein.

Purification The antibody was purified from rabbit antiserum by affinity-chromatography using immunogen.
Purity > 95 %
Antigen
Background

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

Molecular Weight 48 kDa
Gene ID 4609
OMIM 113970
UniProt P01106
Application Notes WB: 1:500~1:1000 IHC: 1:50~1:100 IF: 1:100~1:500 ELISA: 1:40000
Comment

Unigene-Number: Hs.202453 (NCBI Gene Symbol: MYC)

Restrictions For Research Use only
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.
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage -20 °C
Storage Comment Stable at -20°C for at least 1 year.
Expiry Date 12 months
Supplier Images
Western Blotting (WB) image for anti-MYC (AA 31-80) antibody (ABIN1532205) Western blot analysis of extracts from HeLa cells, treated with Forskolin 40nM 30', u...
Immunohistochemistry (IHC) image for anti-MYC (AA 31-80) antibody (ABIN1532205) Immunohistochemistry analysis of paraffin-embedded human breast carcinoma tissue, usi...
Immunofluorescence (IF) image for anti-MYC (AA 31-80) antibody (ABIN1532205) Immunofluorescence analysis of HeLa cells, using MYC (Ab-62) Antibody. The picture on...
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