Strongly staining

XCL1 Synovium RT-PCR (mRNA), ISH (mRNA), IHC Higher levels compared with OA. Expression restricted to lymphocytic infiltrate in RA. Immunostaining shows strong staining in RA compared with OA and reactive arthritis but not PA. 175... 

The conditions for this step have to be set up for every primer by standard tube (in vitro) PCR. The concentration of the different components and the temperature cycle should be the same (or very similar) for both techniques. The number of cycles is lower for in situ PCR, because otherwise one would obtain strong staining that could prevent visualization of the fine morphological details of signal localization. 

Analyze a sample of the most strongly stained cells in the experiment. 

FIGURE 6.7. Immunostained PCNA in the hyperplastic human palatine tonsil, using mouse monoclonal antibody PC10 (diluted 1 10). (A) No heat pretreatment nuclei of the germinal center cells are weakly stained. (B) In contrast, the cells in the same area are strongly stained after simple heating in a hot water bath for 2 hr at 90°C. Reproduced, with permission, from Kawai et al. (1994). Copyright 1994 Blackwell Science Asia. 

The disadvantage of the strong staining properties of phenosafranine and its allied compounds soon led to the discovery of other desensitizers that do not stain to the same extent. Pinakryptol Green was the first to receive wide application. In solution, it has a dark green color and the slight coloration of the film usually disappears with washing. 

An ideal negative cell line control will contain an amount of target antigen, sufficiently low to produce no staining if the procedure has been performed correctly. At the same time, the amount should be sufficiently high to produce a weakly positive stain if the run has been performed under conditions that produce an excessively strong stain. 

An example of the way in which cell line controls can be used is illustrated by Dako s HercepTest kit, which contains three cell line controls a negative, a 1+ (weak staining) and a 3+ (strong staining). All are designed to be placed on the same microscope slide. 

Nonprecipitated proteins that would otherwise be strongly stained must be removed from the gel. The procedure depends on the amount of antiserum present in the gel if antiserum is under 1% (v/v) in the gel, then washing is performed directly (se Step 4 below), when over 1% (v/v), pressing of the gel is necessary. It is performed according to Laurell (4) as follows. 

Each batch of filters should be incubated for the same length of dme. If your second andbody reacts quickly, because of acdvity or the amount bound, then a blue background will appear quickly. However, some do not give a significant background and, therefore, can be left in for 10 min or more to get a strong stain. 

In addition to the four bands of acid phosphatase isoenzyme activity in normal leukocytes, two other isoenzymes may appear in leukemia. In a case of acute granulocytic leukemia with 100% blast forms, only one electrophoretic band of activity was manifest this migrated between normal band 3 and 4 and was therefore designated 3b. In a case of leukemic reticuloendotheliosis with 98% reticulum cells, only one, and this a strongly staining band, was present this migrated anodically beyond No. 4 and was therefore designated No. 5 (L8). 

Bentonite is a crystalline, claylike mineral, and is available as an odorless, pale buff, or cream to grayish-colored fine powder, which is free from grit. It consists of particles about 50-150 pm in size along with numerous particles about 1-2 pm. Microscopic examination of samples stained with alcoholic methylene blue solution reveals strongly stained blue particles. Bentonite may have a slight earthy taste. 

Reveal the enzyme after washing as in step 3, with DAB (Section 17,3.3.1) until a strong staining is obtained (usually 2-5 min), followed by 3 washings as in step 3. 

DBA.44 is a B subset antibody that strongly stains the cytoplasm of most hairy cell leukemias (HCL). However, it is not specific for HCL because it is also... 

A diffuse strong staining with SlOO, negative for CK in a tumor of unknown origin, is good evidence that it may be a melanoma.However, this still needs to be confirmed by additional melanoma markers such as HMB-45, Melan A, tyrosinase, or NKI/C3. This is because... 

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