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ELF1 Antibody - N-terminal region (ARP36845_P050)

100 ul

Regular Price: $289.00

Special Price: $229.00

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Conjugation Options

ARP36845_P050-FITC Conjugated

ARP36845_P050-HRP Conjugated

ARP36845_P050-Biotin Conjugated

Tested Species Reactivity:
Human, Mouse
Predicted Species Reactivity:
Dog, Horse, Human, Mouse, Rabbit, Rat
Product Format:
Liquid. Purified antibody supplied in 1x PBS buffer with 0.09% (w/v) sodium azide and 2% sucrose.
Clonality:
Polyclonal
Host:
Rabbit
Application:
WB
Reconstitution and Storage:
For short term use, store at 2-8C up to 1 week. For long term storage, store at -20C in small aliquots to prevent freeze-thaw cycles.
Replacement Item:
This antibody may replace item sc-133096 from Santa Cruz Biotechnology.
Immunogen:
The immunogen is a synthetic peptide directed towards the N terminal region of mouse ELF1
Purification:
Affinity Purified
Predicted Homology Based on Immunogen Sequence:
Dog: 80%; Horse: 80%; Human: 87%; Mouse: 100%; Rabbit: 87%; Rat: 88%
Complete computational species homology data:
Anti-ELF1 (ARP36845_P050)
Peptide Sequence:
Synthetic peptide located within the following region: DDIVAPITHVSVTLDGIPEVMETQQVQETNADSPGASSPEQRKRKKGRKT
Concentration:
Batch dependent within range: 100 ul at 0.5 - 1 mg/ml
Blocking Peptide:
For anti-ELF1 (ARP36845_P050) antibody is Catalog # AAP36845 (Previous Catalog # AAPP09918)
Datasheets/Manuals:
Printable datasheet for anti-ELF1 (ARP36845_P050) antibody
Target Reference:
Geng,Y., et al., (2005) J. Immunol. 175 (2), 1022-1029
Gene Symbol:
ELF1
Official Gene Full Name:
E74-like factor 1
Alias Symbols:
p70, Sts1, Elf-1, mElf-1
NCBI Gene Id:
13709
Protein Name:
ETS-related transcription factor Elf-1
Description of Target:
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.
Swissprot Id:
Q60775
Protein Accession #:
NP_031946
Nucleotide Accession #:
NM_007920
Protein Size (# AA):
612
Molecular Weight:
67kDa
Tissue Tool:
Find tissues and cell lines supported by DNA array analysis to express ELF1.
RNA Seq:
Find tissues and cell lines supported by RNA-seq analysis to express ELF1.
Protein Interactions:
Ephb4; Dmd; Jak2; Foxp3;

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1 Item(s)

360/12/2019 19:33
  • Overall Experience:
  • Quality:
NYU School of Medicine
  • ARP36845_P050 LOT# QC6776 (Validation done through Antibodies Online Inc)
  • Positive control:  A549 (endogenous ELF1), ELF1 recombinant protein 
  • Negative control:  None 

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 37C 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 50-100ul 1x LDS sample buffer (Life Technologies, B0007). As positive control, take 20µg recombinant ELF1 (Abnova, H00001997P01) 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 (Aviva Systems Biology, ARP36845_P050, 0066160414) diluted 1:500 in TBST containing 5% skim milk ON at 4°C.  o 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  o goat anti-rabbit HRP-conjugated antibody (Invitrogen, G21234) diluted 1:10000 in TBST containing 0.5% skim milk for 1h at RT. o 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 

  • ARP36845_P050 reveals a protein with an apparent molecular weight of the expected MW of 67kDa for ELF1 and some extraneous bands of lower intensity. ARP36845_P050 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. 

Conclusion 

  • Passed. ARP36845_P050 specifically recognizes endogenous ELF1 in A549 cell lysates and recombinant human ELF1.  

Figure legend 

  • Detection of human ELF1 by western blot using ARP36845_P050 diluted 1:500. Samples: whole cell lysate from A549 cells and human ELF1 recombinant protein.
  • Expected molecular weight: approximately 67kDa. 
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