AP31660SU-N PMN antibody

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1 ml / €450.00
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Quick Overview

Rabbit anti Mouse PMN


Product Description for PMN

Rabbit anti Mouse PMN.
Presentation: Serum
Product is tested for Frozen Sections, Cytotoxicity Assay, Paraffin Sections.

Properties for PMN

Product Category Primary Antibodies
Quantity 1 ml
Synonyms Polymorphonuclear Leukocytes
Presentation Serum
Reactivity Ms
Applications C, CT, P
Clonality Polyclonal
Host Rabbit
Shipping to Worldwide
PDF datasheet View Datasheet
Manufacturer Acris Antibodies GmbH
Material safety datasheet MSDS for Polyclonal Antibodies (de)

Datasheet Extract

Application Immunohistochemistry on frozen and paraffin sections.
Cytotoxic assays.
For Cytotoxic Antibodies:
Modified Colormetric Microtiter Assay 1
Results: Antisera of this antibody diluted 1/50 exhibits 80% cytotoxicity on mouse PMN.
Antisera of this antibody diluted 1/50 exhibits <5% cytotoxicity on mouse thymocytes or splenocytes.
For Agglutinating Antibodies:
Antisera dilutions in RPMI-1640 incubated with target cells at 4°C-8°C for 1hr.
Agglutination determined by microscopic observations.
Results: Antisera of this antibody strongly agglutinates mouse PMN but not thymocytes at dilutions to 1/100.
Protocols Rabbit anti-Mouse PMN IHC protocol
Animals were perfused with 10 mM PBS, pH 7.4 followed by 4% paraformaldehyde in 100 mM phosphate buffer, pH 7.4. Tissue was removed and placed in 4% paraformaldehyde for 6 hours at room temperature, then transferred to 10 mM PBS, pH 7.4 in 0.15M isotonic saline at 4°C overnight. Sections were cut at 50 μm on a vibratome using 10 mM PBS and placed in tissue culture wells at 4°C overnight.
1. Pretreat slices in 50 mM NH4Cl (0.267g/100 ml PBS) in PBS at 22°C for 1 hour.
2. Pretreat slices in 0.1% Triton X-100 in PBS at 22°C for 1 hour.
3. Wash in PBS for 5 minutes at 22°C.
4. Block in PBS with 5% NGS at 22°C for 2 hours.
5. Incubate in primary antibody (Rabbit anti-mouse PMN), AP31660SU-N, diluted 1/3000 in PBS with 5% NGS overnight at 4°C.
6. Wash in PBS with 5% NGS at 22°C for 30 minutes.
7. Incubate in secondary antibody, biotinylated goat anti-rabbit IgG in PBS with 5% NGS at 22°C for 1 hour, diluted:
1) 1/200, 125 μl/25 ml
2) Per instructions, Vector ABC Elite Kit, 8 drops/25 ml
8. Wash in PBS with 5% NGS at 22°C for 30 minutes.
9. Block endogenous peroxidase activity. Immediately before use, mix 81 ml PBS, 9 ml methanol and 10 ml 30% H2O2. Incubate for 10 minutes at 22°C.
10. Wash in PBS only 1 x 10 minutes at 22°C.
11. Wash in PBS only 1 x 20 minutes at 22°C
12. Wash in PBS with 5% NGS at 22°C for 30 minutes.
Prepare ABC reagent, if using this option.
13. Incubate in:
1) KPL Streptavidin-peroxidase conjugate diluted 1/200 (125 μl/25 ml) in PBS with 5% NGS, 0.1% Tween 20.
2) Vector ABC elite, diluted according to kit instructions for 1 hour at 22°C.
14. Wash in PBS with 5% NGS at 22°C for 10 minutes.
15. Incubate with DAB (2-5 minutes). Stop reaction with PBS wash.
16. Allow to air dry.
17. Wash salts off in ddH2O. Dehydrate, clear and mount on chromalum gelatin coated slides.
2.76g monobasic
21.45g dibasic heptahydrate
1000 ml ddH2O
40g Paraformaldehyde
PBS (10mM):
0.276 monobasic
2.15g dibasic heptahydrate
100 ml ddH2O
8.76 NaCl/1000 ml
900 ml Isotonic saline + 100 ml mono/dibasic
pH to 7.4
Primary antibody: 17 μl/50 ml PBS with 5% NGS
General Readings
  1. Green et.al., J. Imm. Methods, vol 70: 257, (1984)
Storage Store the antibody at -20°C.
Avoid repeated freezing and thawing.
Shelf life: one year from despatch.
Liquid serum
This antibody reacts to PMN.

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