| CD104 (also GP150 and Integrin beta 4) is a 150-200kD member of the integrin beta family. It forms noncovalent heterodimers with integrin alpha 6 and participates in the formation of epithelial hemidesmosomes. Human integrin beta 4 is a type I transmembrane glycoptotein that is 1795 amino acids (aa) in length. It contains a 683aa extracellular domain (ECD) (aa28-710) and a large 1089aa cytoplasmic region. Two ECD alternate splice forms may exist. One shows an 84aa substitution for the 84aa between aa21-704. A second shows a 144aa substitution for the first 704aa. Human beta 4 ECD shares 88% aa sequence identity with mouse beta4 ECD. |
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| Catalog # | C2446-01H |
| Applications | Suitable for use in Western Blot, Flow Cytometry, Immunocytochemistry and ELISA. Other applications not tested. |
| Recommended Dilutions | Western Blot: 1-2ug/ml; Using a colorimetric detection system, the detection limit for recombinant human Integrin 4 is ~25ng/lane under non-reducing conditions. Reducing conditions are not recommended. Chemiluminescent detection will increase sensitivity by 5-50 fold. |
| Flow Cytometry: 0.2mg/ml; 10ul labels 1-2.5x10e5 human A431 cells in a total reaction volume <200ul. |
| Immunocytochemistry: 10ug/ml detects CD104 in A431 human epidermoid carcinoma cells. Cells were fixed with PBS, 4% paraformaldehyde for 20 minutes at RT and blocked with PBS, 10% normal donkey serum, 0.1% Triton X-100, 1% BSA for 45 minutes at RT. After blocking, cells were incubated with diluted C2446-01H overnight at 4ºC followed by anti-mouse IgG (FITC) at RT in the dark for 1 hour. Cells were washed with PBS, 0.1% BSA between each step. |
| Optimal dilutions to be determined by the researcher. |
| Storage and Stability | Lyophilized powder may be stored at -20°C. Stable for 12 months at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. |
| CAS Number | n/a |
| Clone Type | Monoclonal |
| Isotype | IgG2b |
| Clone No | 7H253 |
| Host | Mouse |
| Source | Human |
| Concentration | ~0.5mg/ml (after reconstitution) |
| Form | Supplied as a lyophilized powder from PBS, 5% trehalose. Reconstitute with 200ul sterile PBS. |
| Purity | Purified by Protein G affinity chromatography. |
| Immunogen | Recombinant protein corresponding to aa28-710 of human Integrin Beta 4 expressed in CHO cells (Accession # P16144). |
| Specificity | Recognizes human CD104 (Integrin beta4). Does not cross react with recombinant human Integrin beta1, beta2, beta3, beta5, beta6, beta8 or recombinant mouse Integrin beta4. |
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| Important Note | This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological. |
