ELF1 antibody (N-Term)

Catalog No. ABIN2779831

The Rabbit Polyclonal anti-ELF1 antibody has been validated for WB and has been independently validated. It is suitable to detect ELF1 in samples from Human, Mouse, Rabbit, Rat, Horse and Dog.

Quick Overview for ELF1 antibody (N-Term) (ABIN2779831)

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

Reactivity: Human, Mouse, Rabbit, Rat, Horse, Dog

Host: Rabbit

Clonality: Polyclonal

Conjugate: This ELF1 antibody is un-conjugated

Application: Western Blotting (WB)

Product Details anti-ELF1 Antibody

Binding Specificity: N-Term

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

Immunogen: The immunogen is a synthetic peptide directed towards the N terminal region of mouse ELF1

Application Details

Application Notes: Optimal working dilutions should be determined experimentally by the investigator.

Comment:

Antigen size: 612 AA

Restrictions: For Research Use only

Independent Validation (WB)

Validation #104222 (Western Blotting)

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

No.: #104222

Date: 10/28/2019

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.

Validation Images

Detection of human ELF1 by western blot using ABIN2779831 diluted 1:500. Samples: whole cell lysate from A549 cells and human ELF1 recombinant protein. Expected molecular weight: approximately 67kDa.

Full Methods

Primary Antibody: ABIN2779831

Secondary Antibody: 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
• • 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.

Handling

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.

Target Details for ELF1

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

Alternative Name: ELF1

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