EIF4E antibody (Eukaryotic Translation Initiation Factor 4E) (AA 170-215)

Details for Product anti-EIF4E Antibody No. ABIN1386772
Antigen
Epitope
AA 170-215
72
15
14
13
11
10
8
6
4
3
3
2
2
1
1
1
1
1
1
1
1
1
Reactivity
Human, Mouse (Murine), Rat (Rattus)
255
121
121
9
9
9
5
5
5
4
3
3
3
Host
Rabbit
210
47
Clonality
Polyclonal
Conjugate
This EIF4E antibody is un-conjugated
15
11
11
8
8
8
6
6
6
6
6
6
2
2
2
2
2
Application
Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Western Blotting (WB)
188
113
110
41
40
35
25
17
15
13
12
5
3
1
1
Options
Immunogen KLH conjugated synthetic peptide derived from human eIF4E
Isotype IgG
Purification Purified by Protein A.
Antigen
Alternative Name Eif4e (EIF4E Antibody Abstract)
Background

Recognizes and binds the 7-methylguanosine-containing mRNA cap during an early step in the initiation of protein synthesis and facilitates ribosome binding by inducing the unwinding of the mRNAs secondary structures. Component of the CYFIP1-EIF4E-FMR1 complex which binds to the mRNA cap and mediates translational repression. In the CYFIP1-EIF4E-FMR1 complex this subunit mediates the binding to the mRNA cap.

Subcellular location: Cytoplasm

Synonyms: CBP, EIF4F, AUTS19, EIF4E1, EIF4EL1, Eukaryotic translation initiation factor 4E, eIF-4E, eIF4E, eIF-4F 25 kDa subunit, mRNA cap-binding protein

Gene ID 1977
UniProt P06730
Application Notes WB 1:100-1000
IHC-P 1:100-500
IF(IHC-P) 1:50-200
IF(ICC) 1:50-200
Restrictions For Research Use only
Format Liquid
Concentration 1 μg/μL
Buffer Aqueous buffered solution containing 1 % BSA, 50 % glycerol and 0.09 % sodium azide.
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
Expiry Date 12 months
Supplier Images
Image no. 1 for anti-Eukaryotic Translation Initiation Factor 4E (EIF4E) (AA 170-215) antibody (ABIN1386772) Image provided by One World Lab validation program. A431 cells probed with Rabbit Ant...
Image no. 2 for anti-Eukaryotic Translation Initiation Factor 4E (EIF4E) (AA 170-215) antibody (ABIN1386772) Formalin-fixed and paraffin embedded human gastric cancer labeled with Anti-eIF4E Pol...
Image no. 3 for anti-Eukaryotic Translation Initiation Factor 4E (EIF4E) (AA 170-215) antibody (ABIN1386772) Independently Validated Antibody, image provided by Science Direct, badge number 0296...
'Independent Validation' Badge
Lot Number 140106
Method validated Immunohistochemistry
Positive Control Human pancreas
Negative Control Human colon
Notes Clear strong signal was detected in positive control sample. Note a minor amount of expected background staining in negative control sample.
Primary Antibody
  • Antigen: Eukaryotic Translation Initiation Factor 4E (EIF4E)
  • Catalog number: ABIN1386772
  • Lot number: 140106
Isotype Control Antibody
  • Antibody: Rabbit IgG isotype control
  • Lot number: C11245
Secondary Antibody
  • Antibody: Biotinylated goat anti-rabbit/anti-mouse (Kit)
  • Lot number: D07640BA
Controls
  • Positive control: Human pancreas tissue stained with antibody
  • Negative control: Human colon tissue stained with antibody
  • Isotype control: Human pancreas tissue stained with isotype control
  • Secondary only control: Human pancreas tissue stained with secondary antibody only
Protocol
  • Immunohistochemistry was performed on a Ventana NEXes automated platform; instrument manufacturer specific reagents are italicized.
  • 1. Slides were preheated in convection oven at 60°C for 30 min
  • 2. Deparaffinization procedure: - 3 changes of Xylene, 5 min each - 3 changes of 100% Ethanol, 3 min each - 3 changes of 95% Ethanol, 3 min each - Rinsed in distilled water, 3 changes
  • 3. Heat retrieval procedure - Slides retrieved in 10.0 mM Citrate, pH6.0 in a 1000W microwave oven (~100°C) for 15 min. - Slides were allowed to cool (in citrate) for 30 min. - Slides were washed x 3 in Distilled water
  • 4. NEXes instrument procedure, iView DAB paraffin protocol (*abridged*): - Slide chamber warmed to 37°C
  • 5. Slides rinsed with *reaction buffer* x3
  • 6. *iView Inhibitor (H2O2)* applied and incubated for 4 min
  • 7. Slides rinsed with *reaction buffer*
  • 8. Antibody Application - Primary antibody diluted 1:250 in PBS (100 microliter applied/slide) - Ventana Isotype control applied neat - Slides Incubated overnight at room temperature (~12 hours ~25°C)
  • 9. Slides rinsed with *reaction buffer* x3
  • 10. *iView Biotinylated IgG* applied and incubated for 8 min
  • 11. Slides rinsed with *reaction buffer*
  • 14. *iView Streptavidin-Horseradish Peroxidase* applied and incubated for 8 min
  • 15. Slides rinsed with *reaction buffer*
  • 16. *iView DAB/H2O2* applied and incubated for 8 min
  • 17. Slides rinsed with *reaction buffer*
  • 18. *iView Copper* applied and incubated for 4 min
  • 19. Slides rinsed with *reaction buffer*
  • 20. Slides washed in Dawn Detergent/tap water
  • 21. Counterstain Procedure - Hematoxylin (Leica 560 MX) 30 sec - Slides washed in tap water, 1 min - Decolorized (10% Acetic Acid in 70% ethanol), 1 min - Slides washed in tap water, 1 min - Bluing (Austin Clear Ammonia), 1 min - Slides washed in tap water, 1 min
  • 22. Dehydration/coverslipping procedure: - 3 changes of 95% Ethanol, 3 min each - 3 changes of 100% Ethanol, 3 min each - 3 changes of Xylene, 5 min each - Mounted with Permount
  • 23. Imaging: Leica SCN 400F Whole Slide Scanner with Digital Image Hub and Leica Slidepath software
Experimental Notes
  • Deviations from protocol/procedure supplied by manufacturer (attached).
  • Step 1: Heated tissue 60°C for 30 minutes; manufacturer heats for 45 minutes.
  • Step 2: No ethanol wash was performed during deparaffinization; manufacturer includes 1 wash of 80% ethanol for 3 minutes.
  • Step 3.1: Slides were heated for 15 minutes; manufacturer provides a range of 15-20 minutes.
  • Step 3.2: Slides were cooled for 30 minutes; manufacturer cools for 20 minutes.
  • Step 4: Italicized reagents and incubation time are fixed instrument parameters.
  • Step 5: Secondary species-specific serum block not used; manufacturer blocks with 5% normal goat serum for 2 hours.
  • Step 8.1: Antibody diluted in PBS at 1:250; manufacture did not recommend diluent or dilution.
  • Step 8.2.1: Primary antibody incubated at room temperature overnight; manufacturer incubates overnight 4°C with agitation.
Validation Images
Immunohistochemistry validation image for anti-Eukaryotic Translation Initiation Factor 4E (EIF4E) (AA 170-215) antibody (ABIN1386772) Figure 1: Human pancreas tissue stained with anti-EIF4E (brown) and counterstained wi...
Immunohistochemistry validation image for anti-Eukaryotic Translation Initiation Factor 4E (EIF4E) (AA 170-215) antibody (ABIN1386772) Figure 2: Human colon tissue stained with anti-EIF4E (brown) and counterstained with ...
Immunohistochemistry validation image for anti-Eukaryotic Translation Initiation Factor 4E (EIF4E) (AA 170-215) antibody (ABIN1386772) Figure 3: Human pancreas tissue stained with isotype control antibody (brown) and cou...
Immunohistochemistry validation image for anti-Eukaryotic Translation Initiation Factor 4E (EIF4E) (AA 170-215) antibody (ABIN1386772) Figure 4: Human pancreas tissue stained with secondary antibody only (brown) and coun...