Steps of the Western blot analysis Free Essay Example, 1000 words

Steps of the Western blot analysis Definition – define the term â€Å"Western blot analysis† Western blot analysis remains a technique that can be used for identifying certain proteins or antibodies in which proteins become separated by the use of electrophoresis, then become transferred to nitrocellulose, and finally reacted with antibody (Pound 2001, p. 38). 2) Principle of the method – briefly explain the principle of the Western blot: This must include the following; a) separation of protein b) The picking up of protein with antibody. Western blot often involves analysis of proteins from cells. Often, cells become lysed to release protein content in them. These proteins content become separated in relation to their protein size by the use of a gel. Furthermore, the separated gel proteins become transferred onto a membrane surface by use of electricity. It becomes the purpose of the membrane to be used as a probe to separate proteins of interest. Often, a primary antibody becomes directed towards the protein of interest and binds to it leaving the nonspecific proteins unbound (Pound 2001, p. 39). Then, a secondary antibody becomes directed to recognize the primary antibody thereby forming a protein-antibody-antibody-sand witch. We will write a custom essay sample on Steps of the Western blot analysis or any topic specifically for you Only $17.96 $11.86/pageorder now Usually, the secondary antibody contain peroxidase enzyme that often converts luminal substrate to give a light releasing substrate. It hence becomes the light which becomes detected as a spot on the film. It becomes the spot size that determines how much protein remained in the spot, in relation to the size of other spots (Michael 2011, p. 56). 2) Steps of the Western blot analysis. List the steps and briefly describe/explain each of them. The first step in western blot analysis remains as the lysis of the cell. Here, the cell becomes lysed with a lysic buffer which contains detergents responsible for breaking the cell’s membrane. In addition, the lysic buffer also contains an inhibitor of the photolytic enzymes which prevent enzymatic degradation of proteins (Rosenberg 2005, p. 66). The second step in western blot analysis involves temperature denaturation. In this step, a sample of protein becomes left to boil for five minutes at a temperature of 95 C. The purpose of that often became to allow protein denaturation to occur. The first step and second step sometimes become referred to as sample preparation steps since they involve preparation of the given proteins to be used in the third step (Rosenberg 2005, p. 66). The third step entails gel electrophoresis.

Enablers of Non-Sequential Internationalization Process among Small Article

Essays on Enablers of Non-Sequential Internationalization Process among Small and Medium-Sized Firms by Osarenkhoe Article The paper "Enablers of Non-Sequential Internationalization Process among Small and Medium-Sized Firms by Osarenkhoe" is a delightful example of an article on marketing. The article entitled â€Å"A study of the enablers of non-sequential internationalization process among small and medium-sized firms† was written by Aihie Osarenkhoe of the University of Gavle (Sweden) Department of Business Studies, and was published in the International Journal of Business Science and Applied Management in February 2008. The objective of the article is to detail the researcher’s findings in a study of internationalization processes followed by small- and medium enterprises. Osarenkhoe’s hypothesis is that firms that do not follow a sequential process towards internationalization are not solely enabled in their non-sequential approach by single factors, as is commonly asserted by the body of prior research, but rather by a number of interconnected factors.The article begins with a basic definition of internationalization – â€Å"the process of increasing involvement in international operations across borders (Welch Luostarinen, 1988)† – and briefly introduces some of the large body of research done in North America and Europe. Both streams of research have, according to Osarenkhoe, developed essentially the same basic framework for models of internationalization:â€Å"Both research traditions conceptualize export development as taking place in gradual and sequential stages (learning sequences involving feedback loops), based on a series of incremental commitment decisions depending on perceptions, expectations, experience, managerial capacity, etc. The firm is assumed to build a stable domestic position before starting international activities.† (p. 2)In other words, internationalization is a step-by-step process undertaken once a firm establishes a sound position in its domestic market; moving into foreign markets is done incre mentally, with each step providing learning to support the following step. The research question of the author then has two elements: First, not all firms follow a sequential, or step-by-step process; and second, their reason for not doing so cannot be attributed to a single factor.The theoretical framework of the research is based on three models that describe sequential internationalization processes. The first is the Uppsala Internationalisation Model, proposed by Johanson and Wiedersheim-Paul in 1975, and expanded by Johanson and Vahlne in 1977. (p. 3) The second is the Transaction Cost Approach (TCA) first proposed by Coase, 1939 and applied to internationalization by Williamson in 1975 (it should be noted, however, that the reference to this latter work is missing from the article).   The third model and most recently-developed model is the Network Approach model, which has been studied by numerous researchers. Osarenkhoe summarises these three models in a table (p. 4).

Derailed †Creative Writing Free Essays

It had rained the night before, and mist was creeping in, like a soft blanket of cold steam. It was silent. The grim reaper was waiting silently for blood. We will write a custom essay sample on Derailed – Creative Writing or any similar topic only for you Order Now Not even the birds dare tweet. The reaper lifts his hefty but swift scythe. Just as he is about to drop it†¦ â€Å"Don’t you worry love, you’re only dreaming again.† Margaret slides out of bed and plods off to fetch a glass of water for Daniel. She stared through the windowpane, and out of the four walls that she and Daniel call home. It was cold and wet, perfect weather for Ducks though. It had rained the night before, and mist was creeping in, like a soft blanket of cold steam. Maggie (as she likes to be called), twisted the cold steel, and a cold transparent liquid flowed out, making a racket as it hit the bottom of the sink. After filling her glass, she shut off the flow of water, and turned sharply around. Just as she did so, she nearly had the shock of her life. In front of her was a hairy ape, stamping towards her with half-closed eyes!! Maggie then put her arms around this beast, and told him to drink the glass of water and to get a bath. Maggie then turned on the TV and was pleased to see that her favourite quiz show was due on in about 15 minutes. Daniel smiled and turned to face the towering stairs. He started to climb the stairs. He reached the landing and for some unknown reason decided to turn around and stare out of the window. He stopped tipped his head to one side and thought about something for a second, said a brief de-ja-vous and carried on with his quest which was to have a bath. Daniel reached the bathroom and started to run a bath. After a brief shave, his bath was ready to dive in to. He dived in and while washing himself, (and doing other bathroom activities) he thought about the day ahead. First activitie of the day was to pack some clothes as he had train journey to deal with and then he would†¦Ã¢â‚¬ ¦Ã¢â‚¬ ¦ Oh, I am so sorry, how can I write a story without describing the main character, and his family to you first. Daniel is a 36-year-old entrepreneur , who, at the age of twenty-two, decided to start his own business , making computer components. He had a bad turn at the age of twenty-six and nearly went bankrupt due to a downfall in the price of shares in the computer components. Now he is doing fine (nearly a millionaire) and him and his wife are very happy and content. He is 5ft 6in and has a distinctive chubby look about him. Daniel has a warm chubby face and sparkling hazel-brown eyes. Handsome for a 36 year old, the females might say. Mggi4e was Daniels sectary until 10 years ago, when they got engaged. Unfortunately, however, she fell on a slippery supermarket floor and nearly broke her neck. She can walk but excessive movement could severely injure her. Maggie is just over 5ft 4in and quite thin. She used to have a wide stature and used to be slightly overweight. She lost this weight due to a strict diet and living regime. Her blonde curly locks hang over her shoulders and are complemented by her warm blue eyes. Anyway, continuing on with what the day held for him:- After his morning shift, Daniel was to call a workers meeting, to outline his plans and to hand out raises, awards ect. Daniel then had the rest of the day off so that he could sleep to prepare to travel down to London to visit his old mate Joe. He would travel by train, which it cheep an if on time quicker than car. It cost a tidy forty pounds and would take 3 hours (from Durham). Not bad and especially for Dan as he would travel by first class. This meant that could enjoy his journey and have drinks, food etc as well. Monday morning (12th May) A shrill ringing woke Dan a 6am. He was staying for two days and started packing accordingly. He didn’t take much as he only needed clothes and deodorant to take with him. He had a few get-well cards from friends, by these didn’t way much and weren’t at all bulky. He then had his break fast and wash etc. Daniel kissed Margaret good by, opened the front door and his garage. A gentle breeze came out of nowhere and I take away card fluttered past and landed at the front of Daniel’s short drive. He dawdled over and picked it up. He looked at it and saw nothing unusual other than a line saying 21:00 no 1365. Dan though nothing of it, only a bit of scrap paper, and got in his car. As he turned the ignition key and placed his luggage in the passenger seat, he felt a weird buttelfy feeling in his stomach. This was odd. Since dan had not exercised properly, he didn’t have any problems. A thought raced across the vast screen of his mind. It might be a stomach ulcer. His had a history of them, but Daniel had been lucky for the past sixteen years. The thought diminished and he reversed his car and carried on. It was only a twenty-minute journey and it passed away quickly and with no problems. The car pulled up at the car park and slowly screeched to a halt. Daniel slid out, paid his two pay and display charge and jumped back in his car. He then practiced the difficult not so accent art of parking and slowly got out. The station was normal. The hustle and bustle of every day lime. Many of the passengers are businessmen and women probably going to work. Dan glanced at his watch, which said 8:30 am. There was time for a coffee. Just as he turned around to head for the cafà ¯Ã‚ ¿Ã‚ ½, a high-pitched squeal dived out of the speakers and into the maze that was Dan’s ears. The squealing halted and the tannoy sounded. â€Å"Could all passengers for the train number 1365 Durham to London please listen. The train will not be here at 21:00 as scheduled but will be delayed for about 15 minutes. These delays are due uncontrollable circumstances. Thank you†. Daniel sighed, as this was his train. All of a sudden, his spine tingled and he sensed a feeling of de-ja-vous. He had 2100 no 1365 on a piece of paper. He could not remember when or where but he knew he had seen it. Dan turned back towards the cafà ¯Ã‚ ¿Ã‚ ½ and walked towards it. Unusually, he felt he was being watched. He strained his neck to glance over his shoulder when a small hand touched his shoulder. Dan jumped and turned sharply around. â€Å"How are you Dan†. It was Joann; Daniel’s secretary from work. Dan replied with a mumbled, â€Å"I’m alright â€Å". Joanne sensed that something was wrong, but thought it best not to ask what. She walked on and faded into the crowd. Dan turned towards the cafà ¯Ã‚ ¿Ã‚ ½ and strolled along towards it with his small luggage, slung over his shoulder. He finally reached his destination and opened the door. Swinging his bag from his shoulder, he sat down. His arse landed on the soft, warm seat (probably warmed by some clean dude who farted constantly) and Dan started to relax. He had hardly been there for a few seconds when an attractive young waiter approached Daniel. She was about 5ft tall and had long locks of curly hair Dangling from her head and down onto her peninsular-like shoulders. She was thin but obviously liked working out, as I could tell from the clear lines on her long arms. Her hands were small and boney, but were terribly scared. The waiter spoke with a small but kind voice. â€Å"Would you like anything sir†, she squeaked. After a few seconds of deliberation, Daniel replied, â€Å"A black coffee please love†. Rachel (it said on her nametag) then chirped â€Å"Be with you in a minute sir†, and then scarpered off to prepare a coffee for Daniel. Just as she said, she returned in less than two minutes with a black coffee, hot and steaming, in a large mug. Daniel sprinkled two sachets of sugar into his mug, and then sipped. He looked around the area where he sat, and sniffed the polluted air. The cafà ¯Ã‚ ¿Ã‚ ½ was small, with the tables in rows of four. The tables had four chairs each and bottles of sauce and condiment sets sat proudly in the middle on top of a tablecloth. The cloth was opulent and excreted a clean, lemon fresh odour that made you think of spring. The cafà ¯Ã‚ ¿Ã‚ ½ was quite eventful and obviously made a good, if not excellent turnover. Daniel finished his coffee and dawdled up to the till to pay. He paid and on his way out, placed a tip of à ¯Ã‚ ¿Ã‚ ½5 in the hand of the waiter. â€Å"Thanks love. Here, have this as a token of my appreciation for good service.† The waiter thanked Daniel and he continued on. All of a sudden the tannoy sounded, following that same annoying squeal. â€Å"Would all passengers for the no 1365 train please make their way to platform four. We are terribly sorry for the delay. Thank you.† Daniel grabbed his bags, and jogged straight up to platform four. To everyone’s bewilderment four police officers, accompanied by four dogs and an armed swat team were standing by the train, carefully looking at everyone suspiciously. The officers and dogs then started to weave in and between passengers, as though they were sniffing for drugs and other illegal objects. The dog came up to Daniel, sniffed his bags and then barked viciously at Daniel. Two officers immediately sprinted over and searched ferociously through his bags. Sure enough, there were drugs in Daniels bag, but they were legal and prescribed by Dan’s doctor. The police officers apologised, helped Daniel to fill his bag, and the toddled over to their positions. The next obstruction was a portable metal detector. Everyone was strolling through it, speculating at what caused this annoying chaos. Dan strolled through it, just as everyone had, and then boarded the train. He got on the train, and stared down the aisle. He glimpsed the door to the first class lounge, and wandered down towards it. The first class lounge was spacious and well ventilated. The buffet counter was in the next carriage to the first class and smelt of common aromas such as pie and chips. The carriage had a slight stench of chip fat and an eerie glimmer of flickering light shed out of the broken bar light. Stepping back into the first class lounge, he looked at his ticket and observed the seat number 321. Daniel walked down the first class lounge, selected his seat and sat on it. He was tired and immediately fell asleep, never to wake again. The train was derailed and no one knows why. The press speculate about why the train was derailed and why there were armed guards at the station. The police won’t let on either. How to cite Derailed – Creative Writing, Papers