Application

Immunoprecipitation from labeled cell extracts: 5 µL of serum is sufficient to precipitate alpha6beta4 from 1-2 x 10E6 cells. In some cases immunoprecipitation from 35S methionine labeled cells results in two bands of 95 and 55 kDa of unknown origin in addition to the 200 kDa and 140 kDa bands corresponding to beta4 and alpha6.

Immunohistochemistry: 5 µg/mL. See protocol below.

Optimal working dilutions must be determined by the end user.



Suggested Protocol for Immunohistochemical:

1. Fix cryostat sections with a 1:1 chloroform-acetone mixture for 10 minutes at room temperature.

2. Air Dry.

3. Wash with 2 mL of 1x PBS.

4. Add 200 µL of first antibody (5 µg/mL) to each slide. Incubate for 1 hour at room temperature in a humidity chamber.

5. Wash with 5 mL of 1x PBS.

6. Add appropriately diluted anti-rabbit secondary antibody conjugate and incubate for 1 hour in a humidity chamber

7. Wash with 5 mL of 1x PBS.

8. Add 200 µL of development solution (containing DAB or other substrate depending on the secondary antibody) and incubate at room temperature for the necessary time to develop the dark color. Follow the reaction with a microscope, to establish the correct color intensity.

9. Block the reaction by washing with 2 mL of 1x PBS.

10. Counterstain with hematoxylin as desired.

Anti-Integrin β4 Antibody is an antibody against Integrin β4 for use in IP, IH.

Immunogen

Synthetic peptide derived from the COOH terminal sequence (cytoplasmic domain) of the human β4 integrin subunit (Tamura et al. 1990). The sequence used is NH2-(K)GTLSTHMDQQFFQT-amide. The lysine (K) is not in the β4 sequence and was added to conjugate the peptid

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Specificity

Specific for beta4 integrin as determined by immunoprecipitation. The epitope recognized is on the cytoplasmic domain of the protein and is thus intracellular. No cross-reactivity to other integrin b subunits.