ELF1 Antikörper (C-Term)

Produktnummer ABIN184790

Der Ziege Polyklonal Anti-ELF1-Antikörper wurde für WB, ELISA und IHC validiert und wurde unabhängig validiert. Er ist geeignet, ELF1 in Proben von Human und Maus zu detektieren.

Kurzübersicht für ELF1 Antikörper (C-Term) (ABIN184790)

Target: ELF1 (E74-Like Factor 1 (Ets Domain Transcription Factor) (ELF1))

Reaktivität: Human, Maus

Wirt: Ziege

Klonalität: Polyklonal

Konjugat: Dieser ELF1 Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), ELISA, Immunohistochemistry (IHC)

Güteklasse: Verified

Produktdetails anti-ELF1 Antikörper

Bindungsspezifität: C-Term

Verwendungszweck: ELF1

Sequenz: AMKQNELLEP NSF

Spezifität: This antibody is expected to recognise both reported isoforms.

Kreuzreaktivität: Human, Maus

Aufreinigung: Purified from goat serum by ammonium sulphate precipitation followed by antigen affinity chromatography using the immunizing peptide.

Immunogen: Peptide with sequence C-AMKQNELLEPNSF, from the C Terminus of the protein sequence according to NP_758961.1, NP_001138825.1.

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise:

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.

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (WB)

Validierung #103876 (Western Blotting)

by: Dittmann Lab, Microbiology Department, NYU Langone Health, NYU School of Medicine

No.: #103876

Datum: 28.10.2019

Antigen: ELF1

Chargennummer: C2P2

Validierte Anwendung: Western Blotting

Positivkontrolle:

A549 (endogenous ELF1), ELF1 recombinant protein

Bewertung:

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

Validierungsbilder

Detection of human ELF1 by western blot using ABIN184790 diluted 1:2500. Samples: whole cell lysate from A549 cells and human ELF1 recombinant protein. Expected molecular weight: approximately 65kDa.

Protokoll

Primärantikörper: ABIN184790

Sekundärantikörper: 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).

Anmerkungen:

• 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.

Handhabung

Format: Liquid

Konzentration: 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.

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Handhabung: Minimize freezing and thawing.

Lagerung: -20 °C

Informationen zur Lagerung: 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.

Details zu ELF1

Target: ELF1 (E74-Like Factor 1 (Ets Domain Transcription Factor) (ELF1))

Andere Bezeichnung: ELF1

Hintergrund: ELF1, E74-like factor 1 (ets domain transcription factor), OTTHUMP00000018310, ets-related transcription factor Elf-1

Gen-ID: 1997, 13709

NCBI Accession: NP_758961, NP_001138825