Immunofluorescence analysis of Metadherin was done on 70% confluent log phase A549 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Metadherin Rabbit Polyclonal Antibody (406500) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d is a merged image showing cytoplasmic localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Dog, Human, Mouse, Non-human primate|
|Published species reactivity||Mouse, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide derived from an internal region of the human, chimpanzee, mouse, and bovine metadherin (Mtdh, metastasis adhesion protein, LYsine-RIch CEACAM1 co-isolated (LYRIC), LYRIC/3D3, astrocyte elevated gene-1 (AEG-1)) protein, which differs from rat by one amino acid|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4|
|Contains||0.1% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/million cells|
|Immunocytochemistry (ICC)||1-2 µg/ml|
|Immunofluorescence (IF)||1-2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Immunoprecipitation (IP)||Assay Dependent|
|Western Blot (WB)||1-3 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
Metadherin (Metastasis adhesion protein), also known as MTDH, LYsine-RIch CEACAM1 co-isolated (LYRIC), is a novel protein that localizes with the tight junction proteins ZO-1 and occludin in polarized epithelial cells. At the tight junction, it acts not as a structural component, but is rather recruited during the maturation of the tight junction complex. Metadherin is overexpressed in breast cancer tissue and breast tumor xenografts, while much lower levels are expressed in normal breast tissue. Metadherin binds to lung vasculature, one of the four common sites of breast cancer metastasis, through a C-terminal segment in the extracellular domain; blocking this lung-homing domain with antibodies or inhibiting metadherin with siRNA has been reported to inhibit breast cancer metastasis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Genetic Deficiency of Mtdh Gene in Mice Causes Male Infertility via Impaired Spermatogenesis and Alterations in the Expression of Small Non-coding RNAs.
40-6500 was used in western blot to examine how metadherin contributes to drug resistance, metastasis, and angiogenesis.
|Meng X,Yang S,Zhang Y,Wang X,Goodfellow RX,Jia Y,Thiel KW,Reyes HD,Yang B,Leslie KK||The Journal of biological chemistry (290:11853)||2015|
Metadherin contributes to the pathogenesis of chronic lymphocytic leukemia partially through Wnt/ß-catenin pathway.
40-6500 was used in western blot to investigate metadherin expression, significance, and related mechanism in chronic lymphocytic leukemia
|Li PP,Feng LL,Chen N,Ge XL,Lv X,Lu K,Ding M,Yuan D,Wang X||Medical oncology (Northwood, London, England) (32:null)||2015|
|Human||1 µg/ml||Exposure to cadmium chloride influences astrocyte-elevated gene-1 (AEG-1) expression in MDA-MB231 human breast cancer cells.||Luparello C,Longo A,Vetrano M||Biochimie (94:207)||2012|