Immunofluorescence analysis of CD30 / TNFRSF8 was done on 70% confluent log phase RAW 264.7 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 of CD30 / TNFRSF8 (HRS4) Mouse Monoclonal Antibody (MA512532) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) 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 and membranous localization. Panel e is a no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Recombinant extracellular domain of CD30 protein|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:40-1:80|
* 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.
MA5-12532 targets CD30 in IHC (P) applications and shows reactivity with Human samples.
The MA5-12532 immunogen is recombinant extracellular domain of CD30 protein.
CD30, a single chain glycoprotein, is synthesized as a 90kDa precursor which is processed in the Golgi complex into a membrane-bound phosphorylated mature 105/120kDa glycoprotein. The CD30/Ki-1 antigen is expressed by mononuclear Hodgkin and multinucleated Reed-Sternberg cells in Hodgkin's disease, by the tumor cells of a majority of anaplastic large cell lymphomas, and by a varying proportion of activated T and B cells. About one third of the Ki-1 positive lymphomas lack the leukocyte common antigen (CD45).
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