ELF1 antibody (C-Term)

Catalog No. ABIN184790

The Goat Polyclonal anti-ELF1 antibody has been validated for WB, ELISA and IHC and has been independently validated. It is suitable to detect ELF1 in samples from Human and Mouse.

Quick Overview for ELF1 antibody (C-Term) (ABIN184790)

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

Reactivity: Human, Mouse

Host: Goat

Clonality: Polyclonal

Conjugate: This ELF1 antibody is un-conjugated

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

Grade: Verified

Product Details anti-ELF1 Antibody

Binding Specificity: C-Term

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.

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 Details

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

Independent Validation (WB)

Validation #103876 (Western Blotting)

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

Notes:

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

Validation Images

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.

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.

Handling

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 Details for ELF1

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

Alternative Name: ELF1

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