CD14 Antibody (M5E2) - BSA Free

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• CD14 Antibody (M5E2)

  Name: Bin Yin

  Application: Immunohistochemistry-Paraffin

  Sample Tested: After bowel and After intestinal bowel

  Species: Grouper

  Verified Customer | Posted 10/16/2020

  This is the graph of the results of the experiment, repeated several times, the results are negative

  Dewaxing to water: 1, paraffin sections in turn sliced into 15 min - dimethylbenzene xylene Ⅰ Ⅱ 15 min - xylene III Ⅰ 5 min - 15 min - anhydrous ethanol anhydrous ethanol Ⅱ 5 min 5 min - 75% - 85% alcohol alcohol 5 min - the distilled water to wash. 2, antigen repair: biopsy under full antigen repair citrate buffer (PH6.0) repair box in a have a certain amount of water pressure cooker, induction cooker heated to stomatal began to jet, stop heating, release the pressure, the slice on the repair box, induction cooker heated to stomatal began to jet, 5 min after close the induction cooker, the process should prevent excessive evaporation buffer, do not dry.After natural cooling, the slides were placed in PBS (PH7.4) and shaken on the decolorizing shaper for 3 times, 5min each. 3. Break endogenous peroxidase: the slides were placed in 3% hydrogen peroxide solution and incubated at room temperature and away from light for 25 min. The slides were placed in PBS (PH7.4) and shaken on a decolorizing shaper for 3 times for 5min each. Serum sealing: 3%BSA was added to cover the tissue evenly in the chemochemical ring, and the tissue was sealed at room temperature for 30min.(Primary antibody is sealed with rabbit serum from goat and BSA from other sources.) 4. Primary antibody was added: the blocking solution was gently removed, the primary antibody prepared with PBS in a certain proportion was added to the sections, and the sections were placed flat in a wet box and incubated overnight at 4°C.(Add a small amount of water in the wet box to prevent evaporation of antibodies) 5. Secondary antibody was added: the slides were placed in PBS (PH7.4) and washed by shaking on the decolorizing shaker for 3 times, 5min each.After the sections were slightly shaken and dried, the tissues were dripped with secondary antibody (HRP-labeled) of the species corresponding to the primary antibody and incubated at room temperature for 50min. 7. DAB color development: The slides were placed in PBS (PH7.4) and shaken on the decoloring shaker for 3 times, 5min each.DAB color developing solution newly prepared was added in the circle after the sections were slightly dried. The color developing time was controlled under the microscope. The positive color was brownish yellow, and the color developing was stopped after the sections were washed under tap water. 8. Restaining nuclei: restaining with hematoxylin for about 3min; washing with tap water; differentiation with hematoxylin differentiation solution for several seconds; washing with tap water; flushing with hematoxylin returning blue solution; washing with running water., dehydration and 9: the slice in turn into 5 min 5 min 75% alcohol - 85% alcohol, anhydrous ethanol Ⅰ Ⅱ 5 min - the anhydrous ethanol dehydration in 5 min - xylene Ⅰ 5 min is transparent, the slice out of xylene is a bit dry, neutral rubber sealing piece. Microscope examination, image acquisition and analysis.

  
  Bio-Techne Response

  This review was submitted through the legacy Novus Innovators Program, reflecting a new species or application tested on a primary antibody.

• CD14 Antibody (M5E2)

  Name: Anushka George-Karote

  Application: Flow Cytometry

  Sample Tested: Whole blood cells

  Species: Canine

  Verified Customer | Posted 05/18/2018

  Monocyte detetction in dog cells using CD14

  The whole blood cells were lysed to get rid of the RBC. The washed cell were then incubated with the Antibody. They were washed and then incubated with a conjugated secondary. Allowed to sit for 20mins and then washed again and read on the BD Fortessa.

  

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