Hank’s balance salt solution

HBBS Hank s balanced salt solution HCA Hypertonic citrate H-CAM Hyaluronic acid cell adhesion molecule HDC Histidine decarboxylase... 

Hank s balanced salt solution (Gibco-BRL, Grand Island, NY). Phosphate-buffered saline ([PBS] Gibco-BRL). 

Add Hank s balanced salt solution (room temperature) to the mixture to bring the vol to 50 mL, and centrifuge the cells for 10 min at 500g. 

Decant the supernatant, and wash the cells with Hank s balanced salt solution two more times. The cells should be relatively free of contaminating red blood cells and platelets after these three washing steps. 

Hank s Balanced Salt Solution (HBSS), pH 7.4 (Gibco, Paisley, Scotland). Store at 15-30°C... 

The suspensions are centrifuged and washed twice in 3% FBS/PBS. They are again centrifuged and washed twice in 3% FBS in PBS with 0.1% Triton X-100. The pellets are resuspended and incubated with 0.5 ml of 1% FBS containing propidium iodide (10 pg/ml Calbiochem, San Diego, CA) and RNAse (1 mg/ml) in Hank s balanced salt solution (without phenol red) for 30 min at 37°C. The samples are analyzed on a Coulter Electronics XL-MCL or a Becton Dickinson FACScan flow cytometer with standard argon ion laser excitation and filter configuration for the FTTC/propidium iodide dye combination. 

Hank s balanced salt solution (HBSS JRH Biosciences). 

Tiypsin-ethylenediamine tetraacetic acid (EDTA) in Hank s balanced salt solution, without calcium or magnesium (Invitrogen, Carlsbad, CA). 

Metastasis 2. Medium for tumor cell culture, e.g., Minimal Essential Medium Eagle (MEM) (Gibco, Invitrogen) 3. 10X Trypsin/EDTA dilute to final concentration lx with PBS (Sigma-Aldrich) 4. Standardized tumor cell suspension, radiolabeled 5. Hank s balanced salt solution, Ca++ and Mg++ free (CMF-HBSS) (Sigma-Aldrich, H-2387) 6. Trypan blue stain 7. Mouse vice 8. Heat lamps... 

Metastasis 2. Hank s balanced salt solution, Ca++ and Mg++ free (CMF-HBSS) 3. Methoxyfluorane or xylazine/ketamine 4. Nail and hair removal cream... 

Prepare daily HBSS/BSA 10 mL sterile 10X Hank s balanced salt solution (HBSS, with phenol red, without Ca2+/Mg2+ Life Technologies, Bethesda, MD),... 

Cells grown to subconfluence are detached by EDTA or enzymatic treatment, appropriately rinsed by centrifugation, and resuspended in Hank s Balanced Salt Solution (HBSS) containing CFDA at 40-100 Aig/ml. After 30 min of incubation at 37°C with gentle agitation, they are rinsed twice in HBSS and once in complete culture medium, and finally plated in complete culture medium in co-culture with unlabeled test cells for the evaluation of gap junction formation. Examination of the cells under the fluorescence microscope to evaluate transfer of the fluorescent dye can be started after 1 h of incubation at 37°C. 

Figures 8.1 and 8.2 show intradermal and mammary fat pad injections, respectively. For photographic purposes, the animals were also sedated for intradermal injections. Both types of injection used 27 gauge needles attached to tuberculin 1-cc syringes. The singlecell suspension was prepared in ice-cold Hank s Balanced Salt Solution (however, any isotonic hquid will work as long as it does not contain serum). Note that the bevel faces the syringe markings, making it easy to see injection volume. Also note that for all of the injections the bevel is oriented so that the bore is visible from the top.

Add Hank s balanced salt solution (room temperature) to the mixture to bring the vol to 50 mL, and centrifuge the cells for 10 min at 500g. Decant the supernatant, and wash the cells with Hank s balanced salt solution two more times. The cells should be relatively free of contaminating red blood cells and platelets after these three washing steps. Perform a cell count. Final cell suspensions should contain >98% granulocytes. 

Hank s Balanced Salt Solution (HBSS) (Mediatech, Inc.). 

Then, cells are detached using 0.25% trypsin-2 mM EDTA in HBSS (Hank s Balanced Salt Solution) and incubated at 55°C for 30 min with a solution of 2 N NaOH to lyses. 

The cerebral cortices of six rats were dissected and immediately placed in ice-cold Hank s balanced salt solution (HBSS) at pH7.5, supplemented with 50 qL/mL penicillin, 50 qg/mL streptomycin and 10 000 KIU/L aprotinin. The cortices were then rinsed twice with ice-cold HBSS and homogenized in 10 vol. of HBSS. The suspension was centrifuged at 2600 rev./min for 10 min at 4°C in a Hitachi CF7D2 centrifuge, and the pellet was homogenized in a buffer... 

Evaluation of the degree of internalization of the radioconjugate at 3 and 4 h was carried out in acidic conditions in order to remove the radioactivity bound to the membrane [16.7]. Labelled DOTATATE that was bound to the membrane but not internalized was removed by incubation of the cells in 20mM sodium acetate in Hank s balanced salt solution (HBSS-Ac) at pH5 for 10 min, followed by centrifugation. The supernatant was collected and the process was repeated once more. The pellet containing the cells was redissolved in 1 mL of O.IN sodium hydroxide, and the radioactivity was measured. 

To the cells previously washed with internalization media (step (h) of the previous protocol), 1 mL of sodium acetate 20niM in Hank s balanced salt solution (HBSS) is added, pH5.0 (see Table I.l for the composition of HBSS). It is also possible to use O.IM glycine buffer at pH2.8. 

TABLE 1.1. COMPOSITION OF HANK S BALANCED SALT SOLUTION (HESS)"... 

No digestion was applied as larvae were allowed to migrate from small boneless muscle pieces to the bottom part (warmed up to 37°C) of a glass reservoir during incubation in Hank s balanced salt solution... 

Caco-2 monolayers are incubated at 37°C for 30 min in a C02 incubator with transport media (TM), prior to initiating a transport experiment. The TM for the apical side is made of Hank s balance salt solution (HBSS) containing lOmM HEPES buffer (pH 7.4) and 25 mM D-glucose (a recent study suggested that simulated intestinal fluid might be a better media to use in the apical compartment especially for highly lipophilic compounds [66,67], and this new media will be investigated in the future). 

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