Immunofluorescence analysis of hnRNP K was performed using 70% confluent log phase HeLa 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 hnRNP K (F45 P9 C7) Mouse Monoclonal Antibody (Product # MA1-087) at 5µg/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) 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 (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear and cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Chicken, Human, Mouse, Rabbit, Rat|
|Host / Isotype||Mouse / IgG1|
|Clone||F45 P9 C7|
|Immunogen||Synthetic peptide, conjugated to Ovalbumin, corresponding to residues S(453) V K Q Y S G K F F(463) of the hnRNP-K protein.|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||5 µg/ml|
|Immunofluorescence (IF)||5 µg/ml|
|Immunohistochemistry (IHC)||5 ug/mL|
|Western Blot (WB)||2 ug/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.
MA1-087 detects the hnRNP-K in human, rat, chicken, rabbit, and mouse cells.
MA1-087 has been successfully used in Western blot, and immunohistochemistry procedures. By Western blot, this antibody detects a ~51 kDa protein.
The MA1-087 immunogen is a synthetic peptide, conjugated to Ovalbumin, corresponding to residues S(453) V K Q Y S G K F F(463) of the hnRNP-K protein.
hnRNP-K (Heterogeneous nuclear ribonucleoprotein K) a component of hnRNP particles, is involved is several steps of gene expression regulation. Human hnRNP-K is a major poly(rC)-binding protein and differs from other hnRNP and quote;s in that nucleic acid binding by hnRNP-K is not mediated by an RNP-CS RNA binding domain.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.