ELF1 antibody (E74-Like Factor 1 (Ets Domain Transcription Factor)) (N-Term)

Details for Product anti-ELF1 Antibody No. ABIN2779831
Target
Binding Specificity
N-Term
12
4
2
2
2
1
1
1
1
1
1
1
Reactivity
Dog, Horse, Human, Mouse, Rabbit, Rat
50
23
20
7
4
2
2
2
1
1
1
1
Host
Rabbit
39
7
4
Clonality
Polyclonal
Conjugate
This ELF1 antibody is un-conjugated
3
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
Application
Western Blotting (WB)
35
18
14
13
7
6
2
1
1
1
1
Options
Immunogen The immunogen is a synthetic peptide directed towards the N terminal region of mouse ELF1
Blocking Peptide Blocking peptide for this product available: ABIN976275
Sequence DDIVAPITHV SVTLDGIPEV METQQVQETN ADSPGASSPE QRKRKKGRKT
Predicted Reactivity Dog: 80%, Horse: 80%, Human: 87%, Mouse: 100%, Rabbit: 87%, Rat: 88%
Characteristics This is a rabbit polyclonal antibody against ELF1. It was validated on Western Blot using a cell lysate as a positive control.
Purification Affinity Purified
Target
Alternative Name ELF1 (ELF1 Antibody Abstract)
Background Elf1 belongs to the ETS family. Elf1 is a transcription factor that activates the LYN and BLK promoters. Elf1 may interact with other transcription factors in order to regulate specific genes. Elf1 can bind to the underphosphorylated form of RB.
Alias Symbols: p70, Sts1, Elf-1, mElf-1
Protein Interaction Partner: Ephb4, Dmd, Jak2, Foxp3,
Protein Size: 612
Molecular Weight 67 kDa
Gene ID 13709
NCBI Accession NM_007920, NP_031946
UniProt Q60775
Application Notes Optimal working dilutions should be determined experimentally by the investigator.
Comment

Antigen size: 612 AA

Restrictions For Research Use only
Format Liquid
Concentration Lot specific
Buffer Liquid. Purified antibody supplied in 1x PBS buffer with 0.09 % (w/v) sodium azide and 2 % sucrose.
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Avoid repeated freeze-thaw cycles.
Storage -20 °C
Storage Comment For short term use, store at 2-8°C up to 1 week. For long term storage, store at -20°C in small aliquots to prevent freeze-thaw cycles.
Images
Image no. 1 for anti-E74-Like Factor 1 (Ets Domain Transcription Factor) (ELF1) (N-Term) antibody (ABIN2779831) WB Suggested Anti-ELF1 Antibody Titration: 0.2-1 ug/ml ELISA Titer: 1:62500 Posit...
'Independent Validation' Badge
Antigen ELF1
Lot Number 66160414
Method validated Western Blotting
Positive Control

A549 (endogenous ELF1), ELF1 recombinant protein

Notes

Passed. ABIN2779831 specifically recognizes endogenous ELF1 in A549 cell lysates and recombinant human ELF1.

Primary Antibody ABIN2779831
Secondary Antibody goat anti-rabbit HRP-conjugated antibody (Invitrogen, G21234)
Protocol
  • Grow A549 cells (ATCC, CCL-185) in DMEM medium (Corning, 10-013-CV) supplemented with 10% FBS (Atlanta biologicals, S11150) and Penicillin-Streptomycin (Corning, 30-002-CI), at 37°C and 5% CO2 to 3x104 cells/cm2 in 2ml on a 6 well plate (Costar, 3516).
  • Wash cells once with 1x PBS treat with trypsin (Corning, 25-053-CI).
  • Take cells up in growth medium.
  • Wash cells once with 1x PBS.
  • Resuspend approximately 2.05x105cells/well in 50-100µl 1x LDS sample buffer (Life Technologies, B0007). As positive control, take 20µg recombinant ELF1 (Abnova, H00001997-P01) in 1x LDS sample buffer.
  • Denature samples for 3min at 95°C and subsequently keep them on ice.
  • Separate samples on a Bolt 4-12% Bis-Tris Plus Gel (Invitrogen, NW04122) in an electrophoresis chamber (Mini Gel Tank, Invitrogen, A25977) for 30min at 80V and then for 60min at 120V.
  • Transfer proteins onto nitrocellulose membrane (Invitrogen, IB23002) using an iBlot 2 Gel Transfer Device (Invitrogen).
  • Block the membrane with TBS containing 0.05% Tween (TBST) containing 5% skim milk for 1h at RT.
  • Incubation with primary
    • o rabbit anti-ELF1 antibody (antibodies-online, ABIN2779831, 0066160414) diluted 1:500 in TBST containing 5% skim milk ON at 4°C.
    • loading control mouse anti-beta actin antibody (Invitrogen, MA5-15739) diluted 1:5000 in TBST containing 0.5% skim milk ON at 4°C.
  • Wash membrane 3x for 10min with TBST.
  • Incubation with secondary
    • goat anti-rabbit HRP-conjugated antibody (Invitrogen, G21234) diluted 1:10000 in TBST containing 0.5% skim milk for 1h at RT.
    • goat anti-mouse HRP-conjugated antibody (Invitrogen, G21040) diluted 1:10000 in TBST containing 0.5% skim milk for 1h at RT.
  • Wash membrane 3x for 10min with TBST.
  • Reveal protein bands using SuperSignal West Dura Extended Duration Substrate (ThermoFisher Scientific, 34075) and a ChemiDoc MP Imaging System (Bio-Rad).
Experimental Notes
  • ABIN2779831 reveals a protein with an apparent molecular weight of the expected MW of 67kDa for ELF1 and some extraneous bands of lower intensity. ABIN2779831 also reveals a recombinant GST-tagged protein at the expected molecular weight (theoretical MW 93.9kDa).

  • Other ELF1-antibody dilutions were tested, but a dilution of 1:5000 was found optimal in terms of minimal background and strength of signal.

Validation Images
Western Blotting validation image for anti-E74-Like Factor 1 (Ets Domain Transcription Factor) (ELF1) (N-Term) antibody (ABIN2779831) Detection of human ELF1 by western blot using ABIN2779831 diluted 1:500. Samples: who...