ELF1 antibody (C-Term)

Catalog No. ABIN6261511

Product Details anti-ELF1 Antibody

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

Binding Specificity: C-Term

Reactivity: Human

Host: Rabbit

Clonality: Polyclonal

Conjugate: This ELF1 antibody is un-conjugated

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

Specificity: ELF1 Antibody detects endogenous levels of total ELF1.

Predicted Reactivity: Horse

Purification: 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.

Isotype: IgG

Application Details

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

Restrictions: For Research Use only

Independent Validation (WB)

Validation #104221 (Western Blotting)

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

No.: #104221

Date: 10/28/2019

Antigen: ELF1

Lot Number: 4318850

Method validated: Western Blotting

Positive Control:

A549 (endogenous ELF1), ELF1 recombinant protein

Notes:

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

Validation Images

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.

Full Methods

Primary Antibody: ABIN6261511

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
• • 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).

Experimental Notes:

• 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.

Handling

Format: Liquid

Concentration: 1 mg/mL

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

Preservative: Sodium azide

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

Storage: -20 °C

Storage Comment: Store at -20 °C. Stable for 12 months from date of receipt.

Expiry Date: 12 months

Target Details for ELF1

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:

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

Molecular Weight: 67kDa

Gene ID: 1997

UniProt: P32519