These are high quality immunohistochemistry human positive control tissues (FFPE) for helicobacter pylori.
Note: Human stomach tissue containing helicobacter pylori bacteria are coccoid in shape characterizing the resting or dormant state. This in no way reduces the antigenicity of the bacteria.
The first slide (1) is stained using Scytek’s Helicobacter Pylori antibody for authenticity before sending to you (see protocol below).
These tissues are designed and specified for IHC, they are not optimal for other stain types. No other staining has been performed other than H&E for verification.
Human tissues are cut at 5um, placed at top third of a positively charged microscope slide. These slides have been used by us for more than 4 years with routine, special, IHC, IF (hand), Dako and other machine types (Ventana) all over the world.
Unstained slides are serially sectioned and air dried overnight, then baked at 60C for at least 1 hour before shipment.
Every stained slide is microscopically checked for positive material and its corresponding box is labeled with a lot number.
Helicobacter Pylori Antibody Protocol
- Run slides to distilled water.
- HIER using sodium citrate pH 6.0 solution in pressure cooker (steam) for 20 minutes.
- Cool with lid off, power off 20 minutes.
- Wash under cold running water, 10 minutes
- Block with 3% hydrogen peroxide – 20 minutes
- Wash – PBST – 2 X 5 minutes
- Block with 2.5% normal horse serum – 20 minutes
- Without rinsing, shake off excess horse serum.
- Primary antibody at 1:100 for at least 45 minutes. Make up in PBST with 2.5% horse serum.
- Wash with PBST 2 X 5 minutes
- Apply secondary antibody, anti-rabbit – 45 minutes
- Wash in PBS 2 X 5 minutes
- Prepare DAB brown (Vector DAB Brown).
- Apply 50-200 ul to positive controls and watch under microscope – record time of positive control. To stop DAB reaction, use distilled water.
- Wash in distilled water 5 minutes.
- Pass Hematoxylin solution through 0.2 mm filter
- Repeatedly dip slides in Hematoxylin for 30 seconds
- Rinse with water by dipping in water bath for 10 seconds
- Submerge in bath of 0.1% sodium bicarbonate solution for 30 seconds
- Rinse with water by dipping in water bath for 5 seconds
- Submerge slides in running water for 5 minutes
- Dehydrate in 2 changes of 95% alcohol, 100% alcohol and clear in 2 changes of xylene.
- Cover slip
All other orders – Current turnaround times for control slides – Cut to order – 1 – 3 weeks.