Question

Cytosol Anti-Dnak IP 1- Dnak ITIL IT | 1 2 3 4 5 6 wt Adnak wt after 1% SDS Adnak / labeled Adnak +unlabeled wt Figure 1. Transient Interaction of Newly Synthesized Proteins with Dnak E. coli spheroplasts were pulse-chase labeled at 30°C, followed by lysis and coimmunoprecipitation (co-IP) of DnaK-polypeptide complexes. (A) SDS-PAGE of Dnak and associated polypeptides. A 15 s pulse of [S]methionine was followed by addition of excess nonradioactive methionine for 15 s, followed by lysis. Total radiolabeled proteins in the soluble fraction (cytosol) of wild-type (MC4100) (lane 1) and Adnak (BB1553) spheroplasts (lane 2) were immunoprecipitated with anti-Dnak antibody (lanes 3 and 5; equivalent to 15 times the amount loaded in lanes 1 and 2). IP was performed following denaturation with SDS and subsequent dilution (lane 4). IP (as in lanes 3 and 5 was performed, but from a 1:1 mixture of radiolabeled Adnak and unlabeled wild-type spheroplasts (lane 6).

1. 1. Briefly describe why an immune precipitation (IP) was used in this figure?
2. 2. Compare lanes 1 and 3?
      1. What question is being asked?

2. What is the result?

I need help with A and B please. can you explain how you got that answer? Its for cell and molecular.

Answer 1

ANSWER:-

S-35 methionine was used to radiolabeled the DNaK as it is a protein consist of amino acids since methioine is a amino acid so it when DNaK will be synthesized it S-35 will be incorporated so it will label specifically protein only.

B. When compared with lane 1and 2 with lane 3 & 5 it shows that DNaK conc is very low bcz in lane 3 & 5 , there is 15 times the amount loaded in loaded in lane 1&2. Aiso in mutant lane there is no DNa-K when detected with anti - DNaK ANTIBODY IN LANE 5.

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