EIF4E antibody (AA 121-217)

Catalog No. ABIN1386772

Product Details anti-EIF4E Antibody

Target: EIF4E (Eukaryotic Translation Initiation Factor 4E (EIF4E))

Binding Specificity: AA 121-217

Reactivity: Human, Rat

Host: Rabbit

Clonality: Polyclonal

Conjugate: This EIF4E antibody is un-conjugated

Application: Western Blotting (WB), ELISA, Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunocytochemistry (ICC), Immunohistochemistry (Frozen Sections) (IHC (fro))

Cross-Reactivity: Human, Rat

Predicted Reactivity: Mouse

Purification: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human eIF4E

Isotype: IgG

Application Details

Application Notes: WB 1:300-5000
ELISA 1:500-1000
IHC-P 1:200-400
IHC-F 1:100-500
IF(IHC-P) 1:50-200
IF(IHC-F) 1:50-200
IF(ICC) 1:50-200
ICC 1:100-500

Restrictions: For Research Use only

Independent Validation (IHC)

Validation #029640 (Immunohistochemistry)

by: Immunohistochemistry Core, NYU Langone

No.: #029640

Date: 03/14/2014

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.

Validation Images

Figure 1: Human pancreas tissue stained with anti-EIF4E (brown) and counterstained with hematoxylin.

Figure 2: Human colon tissue stained with anti-EIF4E (brown) and counterstained with hematoxylin.

Figure 3: Human pancreas tissue stained with isotype control antibody (brown) and counterstained with hematoxylin.

Figure 4: Human pancreas tissue stained with secondary antibody only (brown) and counterstained with hematoxylin.

Full Methods

Primary Antibody:

• Antigen: Eukaryotic Translation Initiation Factor 4E (EIF4E)
• Catalog number: ABIN1386772
• Lot number: 140106

Secondary Antibody:

• Antibody: Biotinylated goat anti-rabbit/anti-mouse (Kit)
• Lot number: D07640BA

Full 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.

Handling

Format: Liquid

Concentration: 1 μg/μL

Buffer: 0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.

Preservative: ProClin

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

Storage: 4 °C,-20 °C

Storage Comment: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Expiry Date: 12 months

Target Details for EIF4E

Target: EIF4E (Eukaryotic Translation Initiation Factor 4E (EIF4E))

Alternative Name: eIF4E (EIF4E Products)

Synonyms: AUTS19 antibody, CBP antibody, EIF4E1 antibody, EIF4EL1 antibody, EIF4F antibody, eif4e-1 antibody, zgc:86680 antibody, EG668879 antibody, Eif4e-ps antibody, If4e antibody, eIF-4E antibody, 0260/09 antibody, 0587/11 antibody, 0589/11 antibody, 0919/12 antibody, 1004/13 antibody, 2 antibody, 7238 antibody, CG4035 antibody, CT13384 antibody, CT39424 antibody, CT39426 antibody, D-eIF4E antibody, Dmel\\CG4035 antibody, Eif4E antibody, Eif4e antibody, IF4E antibody, dEif4e antibody, deIF-4E antibody, deIF4E antibody, eIF 4E antibody, eIF-4E1 antibody, eIF-4E2 antibody, eIF-4EII antibody, eIF-4e antibody, eIF4E antibody, eIF4E-1 antibody, eIF4E-1/2 antibody, eIF4E-2 antibody, eIF4E1 antibody, eIF4EI antibody, eIF4e antibody, eif-4E antibody, eif-4e antibody, eif4e antibody, l(3)07238 antibody, l(3)67Af antibody, l(3)S025007 antibody, l(3)S026009 antibody, l(3)S058711 antibody, l(3)S091912 antibody, cbp antibody, eif4f antibody, eif4e1 antibody, eif4el1 antibody, EIF-4E antibody, zgc:101581 antibody, eukaryotic translation initiation factor 4E antibody, eukaryotic translation initiation factor 4ea antibody, Eukaryotic initiation factor 4E antibody, eucaryotic initiation factor 6 antibody, eukaryotic translation initiation factor 4E L homeolog antibody, eukaryotic translation initiation factor 4E family member 1B S homeolog antibody, translation initiation factor eIF4E antibody, eukaryotic translation initiation factor 4eb antibody, EIF4E antibody, eif4ea antibody, Eif4e antibody, eIF-4E antibody, eif6 antibody, eif4e antibody, eif4e.L antibody, eif4e1b.S antibody, LOC100533326 antibody, eif4eb antibody

Background:

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

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.

Gene ID: 1977

UniProt: P06730

Pathways: BCR Signaling