hi naymul, whatever the images u took i thnk  those are not WB images, did u performed next steps after transfering protein to membrane such as blocking-washing-pri AB overnight incubation 4 degree - washing- Sec AB incubation - washing- then blot development.

after performing these steps if u get the same results then increase blocking buffer concentration as well as incubation time

ur blot as lots of non - soecificity band 

To reiterate Daniel's point, a Ponceau stain will show you all the proteins in your sample, whereas antibody labeling will show a specific protein by labeling a specific antigen. A Ponceau stain is a good precautionary step before antibody labeling to ensure you have achieved adequate and even transfer, and that you loaded approximately equal protein to each of your lanes.

However, a Ponceau stain should show more clear bands throughout the entire membrane, instead of one large smear as seen in your photo. This indicates to me that you may have major protein degradation going on in your samples. Are you using protease inhibitors in your Liver homogenates?

Yes. I have used protease inhibitors

hello lyndsey  please look in to that blot properly, u will get smear band oly because lot of non specific protein will b there in our samples thats y we we  get  compact/smear bands...but in this blot  there is a protein transfer he has to move forward for others steps such as blocking antibody incubation and blot development..then we will get clear picture of this problem ...and i agree with you he has to very careful about protein degradation   

Hi Basavaprabhu, yes I am aware he has to proceed with the actual labeling part of the immunoblot to see any specific proteins (as I stated above). However, after a transfer I typically see some clearly defined bands, especially of cytoskeletal proteins that are in huge abundance. Granted, you can not say for certain what these bands are, as Ponceau stain labels all proteins. However,  in a Ponceau you can usually see a band pretty clearly at around 40 kD that corresponds to the MW of actin. 

Naymul,  my point is this: Yes, go ahead with the labeling as others have suggested. That is the only way you will find out if your protein of interest is present. However, make sure to also label for a loading control like beta-tubulin. The smear in your Ponceau and lack of the clear bands typically seen around 40kD means you might have some protein degradation. Liver cells contain more proteases than other tissue types, and you may need to adjust your protease inhibitor cocktail accordingly (see attachment). I hope this clarifies my point. 

Good luck!

Thank you very much for your valuable suggestion Miss Lyndsey and Mr. Basavaprabhu.I will follow it