D369-3
Page 1For Research Use Only.
Not for use in diagnostic procedures.

Anti-H. pylori mAb
CODE No.

D369-3

CLONALITY
CLONEISOTYPE
QUANTITY

Monoclonal
TMDU-D8
Mouse IgG3 κ100 µL, 1 mg/mL

SOURCEIMMUNOGEN

Purified IgG from hybridoma supernatant
Sonicated whole bacterial lysate of H.pylori (a combination of strains ATCC 43504, ATCC43579, ATCC 43629)
PBS containing 50% Glycerol (pH 7.2). No preservative is contained.
This antibody solution is stable for one year from the date of purchase when stored at -20°C.

FORMULATIONSTORAGEAPPLICATION-CONFIRMED

Immunohistochemistry 1 µg/mL (paraffin section)
Heat treatment for paraffin embedded section: microwave oven, for 40 min. in 10 mM citrate buffer (pH 6.2)

SPECIES CROSS REACTIVITY on IHCSpecies

HumanMouse

Rat

Hamster

SampleH. pylori-infected stomach

Not tested

Not tested

Not tested

Reactivity+REFERENCES

1) Munari, M., et al., Blood. 16, 6612-6616 (2011)
2) Ito, T., et al., Lab. Invest. 88, 664-681 (2008)

For more information, please visit our web site http://ruo.mbl.co.jp/

MEDICAL & BIOLOGICAL LABORATORIES CO., LTD.
URL http://ruo.mbl.co.jp/e-mail support@mbl.co.jp, TEL 052-238-1904

D369-3
Page 2

RELATED PRODUCTS
D369-3
M078-3Anti-H. pylori mAb
Mouse IgG3 (isotype control)D369-3
Page 3

Immunohistochemistry for formalin fixed paraffin-embedded section1) Deparaffinize tissue sections in Xylene 3 times for 5 min. each.
2) Immerse the slides with Ethanol 3 times for 5 min. each.
3) Immerse the slides with PBS 3 times for 5 min.
4) Remove the slides from PBS and heat-treat with 10 mM Citrate buffer (pH 6.2) for 40 min. at 97°C using microwave oven.
5) Let the slide cool down until at room temperature in the Citrate buffer.
6) Remove the slides from the Citrate buffer and inactivate endogenous peroxidase with 3% H2O2 in Methanol for 10 min.
7) Wash the slides with PBS 2 times for 5 min each.
8) Remove the slides from PBS, and immerse the slides in Blocking buffer [20 mM HEPES, 1% BSA, 135 mM NaCl] for 5 min.
at room temperature to block non-specific staining. Do not wash.
9) Tip off the blocking buffer, wipe gently around each section and cover tissues with primary antibody diluted with Blocking
buffer as suggested in the APPLICATION. (The concentration of antibody will depend on the conditions.) Incubate thesections for 1 hr. at room temperature.
10) Wash the slides 2 times in PBS for 5 min. each.
11) Incubate the section with Histostar (Ms + Rb) (MBL; code no. 8460) for 30 min. at room temperature.
12) Wash the slides 3 times in PBS for 5 min. each.
13) Visualize by reacting for 10 min. with Histostar DAB Substrate Solution (MBL; code no. 8469). *DAB is a suspect
carcinogen and must be handled with care. Always wear gloves.
14) Wash the slides 2 times in PBS for 5 min. each.
15) Counterstain in hematoxylin for 5 min., wash the slides 3 times in water for 5 min. each, and then immerse the slides in PBS
for 5 min.
16) Dehydrate by immersing in Ethanol 3 times for 5 min. each, followed by immersing in Xylene 3 times for 3 min. each. Now
ready for mounting.
(Positive control for Immunohistochemistry; H. pylori-infected stomach)

Immunohistochemical detection of H. pylori in human stomach
Left: H. pylori positive case (arrowhead: in mucous layer, arrow: in lamina propria)
Right: H. pylori negative case
Brown: Anti-H. pylori mAb (D369-3)
Blue: Hematoxylin
The data were kindly provided by Prof. Yoshinobu Eishi1 and Mr. Keisuke Uchida2.
(1Department of Human Pathology, Tokyo Medical and Dental University Graduate School,
2
Division of Surgical Pathology, Tokyo Medical and Dental University Hospital).

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