EIF4E Antikörper (AA 121-217)

Produktnummer ABIN1386772

Produktdetails anti-EIF4E Antikörper

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

Bindungsspezifität: AA 121-217

Reaktivität: Human, Ratte

Wirt: Kaninchen

Klonalität: Polyklonal

Konjugat: Dieser EIF4E Antikörper ist unkonjugiert

Applikation: 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))

Kreuzreaktivität: Human, Ratte

Homologie: Mouse

Aufreinigung: Purified by Protein A.

Immunogen: KLH conjugated synthetic peptide derived from human eIF4E

Isotyp: IgG

Anwendungsinformationen

Applikationshinweise: 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

Beschränkungen: Nur für Forschungszwecke einsetzbar

Independent Validation (IHC)

Validierung #029640 (Immunohistochemistry)

by: Immunohistochemistry Core, NYU Langone

No.: #029640

Datum: 14.03.2014

Chargennummer: 140106

Validierte Anwendung: Immunohistochemistry

Positivkontrolle: Human pancreas

Negativkontrolle: Human colon

Bewertung: Clear strong signal was detected in positive control sample. Note a minor amount of expected background staining in negative control sample.

Validierungsbilder

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.

Protokoll

Primärantikörper:

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

Sekundärantikörper:

• 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

Anmerkungen:

• 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.

Handhabung

Format: Liquid

Konzentration: 1 μg/μL

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

Konservierungsmittel: ProClin

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

Lagerung: 4 °C,-20 °C

Informationen zur Lagerung: Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.

Haltbarkeit: 12 months

Details zu EIF4E

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

Andere Bezeichnung: eIF4E (EIF4E Produkte)

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

Hintergrund:

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.

Gen-ID: 1977

UniProt: P06730

Pathways: BCR Signaling