ELF1 Antikörper (C-Term)

Produktnummer ABIN6261511

Dieses Anti-ELF1-Antikörper ist ein Kaninchen Polyklonal-Antikörper zur Detektion von ELF1 in WB, ELISA, IF und ICC. Geeignet für Human. Unabhängig validiert für die Verwendung in Western Blotting.

Kurzübersicht für ELF1 Antikörper (C-Term) (ABIN6261511)

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

Reaktivität: Human

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser ELF1 Antikörper ist unkonjugiert

Applikation: Western Blotting (WB), ELISA, Immunofluorescence (IF), Immunocytochemistry (ICC)

Produktdetails anti-ELF1 Antikörper

Bindungsspezifität: C-Term

Spezifität: ELF1 Antibody detects endogenous levels of total ELF1.

Homologie: Horse

Aufreinigung: The antiserum was purified by peptide affinity chromatography using SulfoLink^TM Coupling Resin (Thermo Fisher Scientific).

Immunogen: A synthesized peptide derived from human ELF1, corresponding to a region within C-terminal amino acids.

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: WB 1:500-1:2000, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (WB)

Validierung #104221 (Western Blotting)

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

No.: #104221

Datum: 28.10.2019

Antigen: ELF1

Chargennummer: 4318850

Validierte Anwendung: Western Blotting

Positivkontrolle:

A549 (endogenous ELF1), ELF1 recombinant protein

Bewertung:

Passed. ABIN6261511 recognizes recombinant and endogenous human ELF1. The antibody does also reveal an additional larger MW protein band in A549 cell lysates.

Validierungsbilder

Detection of human ELF1 by western blot using ABIN6261511 diluted 1:1000. Samples: whole cell lysate from A549 cells and human ELF1 recombinant protein. Expected molecular weight: approximately 67kDa.

Protokoll

Primärantikörper: ABIN6261511

Sekundärantikörper: goat anti-rabbit HRP-conjugated antibody (Invitrogen, G21234)

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
• • rabbit anti-ELF1 antibody (antibodies-online, ABIN6261511, 4318850) diluted 1:1000 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
• • 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).

Anmerkungen:

• ABIN6261511 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: 1 mg/mL

Buffer: Rabbit IgG in phosphate buffered saline , pH 7.4, 150 mM NaCl, 0.02 % sodium azide and 50 % glycerol.

Konservierungsmittel: Sodium azide

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

Lagerung: -20 °C

Informationen zur Lagerung: Store at -20 °C. Stable for 12 months from date of receipt.

Haltbarkeit: 12 months

Details zu ELF1

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

Andere Bezeichnung: ELF1

Hintergrund:

Description: Transcription factor that activates the LYN and BLK promoters. Appears to be required for the T-cell-receptor-mediated trans activation of HIV-2 gene expression. Binds specifically to two purine-rich motifs in the HIV-2 enhancer.

Gene: ELF1

Molekulargewicht: 67kDa

Gen-ID: 1997

UniProt: P32519