EIF4E anticorps (AA 121-217)

N° du produit ABIN1386772

Détail du produit anti-EIF4E anticorps

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

Épitope: AA 121-217

Reactivité: Humain, Rat

Hôte: Lapin

Clonalité: Polyclonal

Conjugué: Cet anticorp EIF4E est non-conjugé

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))

Réactivité croisée: Humain, Rat

Homologie: Mouse

Purification: Purified by Protein A.

Immunogène: KLH conjugated synthetic peptide derived from human eIF4E

Isotype: IgG

Information d'application

Indications d'application: 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: 14.03.2014

Numéro du lot: 140106

Application validée: Immunohistochemistry

Contrôle positif: Human pancreas

Contrôle négative: Human colon

Conclusion: 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.

Protocole

Anticorps primaire:

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

Anticorps secondaire:

• 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

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.

Stockage

Format: Liquid

Concentration: 1 μg/μL

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

Agent conservateur: ProClin

Précaution d'utilisation: This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.

Stock: 4 °C,-20 °C

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

Date de péremption: 12 months

Détails sur EIF4E

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

Autre désignation: eIF4E (EIF4E Produits)

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

Sujet:

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.

ID gène: 1977

UniProt: P06730

Pathways: BCR Signaling