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ELF1 antibody (C-Term)

Verified ELF1 Reactivity: Human, Mouse WB, ELISA, IHC Host: Goat Polyclonal unconjugated
Catalog No. ABIN184790
  • Target See all ELF1 Antibodies
    ELF1 (E74-Like Factor 1 (Ets Domain Transcription Factor) (ELF1))
    Binding Specificity
    • 15
    • 4
    • 4
    • 4
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    C-Term
    Reactivity
    • 39
    • 7
    • 7
    • 6
    • 4
    • 3
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    Human, Mouse
    Host
    • 36
    • 3
    Goat
    Clonality
    • 38
    • 1
    Polyclonal
    Conjugate
    • 25
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    This ELF1 antibody is un-conjugated
    Application
    • 36
    • 13
    • 13
    • 11
    • 10
    • 7
    • 6
    • 3
    • 2
    • 1
    • 1
    • 1
    Western Blotting (WB), ELISA, Immunohistochemistry (IHC)
    Purpose
    ELF1
    Sequence
    AMKQNELLEP NSF
    Specificity
    This antibody is expected to recognise both reported isoforms.
    Cross-Reactivity
    Human, Mouse
    Purification
    Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.
    Grade
    Verified
    Immunogen
    Peptide with sequence C-AMKQNELLEPNSF, from the C Terminus of the protein sequence according to NP_758961.1, NP_001138825.1.
    Isotype
    IgG
  • Application Notes

    Immunohistochemistry: Paraffin embedded Human Thymus. Recommended concentration: 2.5 μg/mL.

    Western Blot: Approx 65 kDa band observed in Human Lymph Node and Tonsil and in Mouse Thymus lysates (calculated MW of 64.9 kDa according to Human NP_001138825.1 and 62.3 kDa according to Mouse NP_001273341.1). Recommended concentration: 0.1-0.3 μg/mL. Appro

    Peptide ELISA: antibody detection limit dilution 1:64000.

    Restrictions
    For Research Use only
  • Validation #103876 (Western Blotting)
    'Independent Validation' Badge
    by
    Dittmann Lab, Microbiology Department, NYU Langone Health, NYU School of Medicine
    No.
    #103876
    Date
    10/28/2019
    Antigen
    ELF1
    Lot Number
    C2P2
    Method validated
    Western Blotting
    Positive Control

    A549 (endogenous ELF1), ELF1 recombinant protein

    Negative Control
    Notes

    Passed. ABIN184790 specifically recognizes endogenous ELF1 in A549 cell lysates and recombinant human ELF1.

    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN184790
    Secondary Antibody
    rabbit anti-goat HRP-conjugated antibody (Invitrogen, 31402)
    Full 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
      • goat anti-ELF1 antibody (antibodies-online, ABIN184790, C2P2) diluted 1:2500 in TBST containing 0.5% 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
      • rabbit anti-goat HRP-conjugated antibody (Invitrogen, 31402) diluted 1:20000 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
    • ABIN184790 reveals a protein with an apparent molecular weight of approximately 62kDa. The expected molecular weight for endogenous human ELF1 is approximately 67kDa. The antibody 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 1:2500 was found optimal in terms of minimal background and strength of signal.

  • Format
    Liquid
    Concentration
    0.5 mg/mL
    Buffer
    Supplied at 0.5 mg/mL in Tris saline, 0.02 % sodium azide, pH 7.3 with 0.5 % bovine serum albumin.
    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
    Minimize freezing and thawing.
    Storage
    -20 °C
    Storage Comment
    Aliquot and store at -20°C, with minimal freeze/thawing. A working aliquot may be refrigerated at 4°C for a few weeks and still remain viable.
  • Target
    ELF1 (E74-Like Factor 1 (Ets Domain Transcription Factor) (ELF1))
    Alternative Name
    ELF1 (ELF1 Products)
    Synonyms
    zef1 antibody, fb05d07 antibody, wu:fb05d07 antibody, Elf-1 antibody, Sts1 antibody, mElf-1 antibody, p70 antibody, xelf1 antibody, elf-1 antibody, E74 like ETS transcription factor 1 antibody, E74-like ETS transcription factor 1 antibody, E74-like factor 1 antibody, E74 like ETS transcription factor 1 L homeolog antibody, E74 like ETS transcription factor 1 S homeolog antibody, ELF1 antibody, elf1 antibody, Elf1 antibody, elf1.L antibody, elf1.S antibody
    Background
    ELF1, E74-like factor 1 (ets domain transcription factor), OTTHUMP00000018310, ets-related transcription factor Elf-1
    Gene ID
    1997, 13709
    NCBI Accession
    NP_758961, NP_001138825
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