For Research Use Only.
Not for use in diagnostic procedures.

PD029
Lot 012~Page 1POLYCLONAL ANTIBODY

Anti-Sendai Virus pAb
Code No.
PD029

Quantity50 L

Form
Serum

BACKGROUND: Sendai virus (SeV), also known as

APPLICATIONS:

murine parainfluenza virus type 1, is a negative sense,
single-stranded RNA virus of the paramyxovirus subfamily
Paramyxovirinae, genus Respirovirus, members of whichprimarily infect mammals. It was originally recovered inSendai, Japan, so it is also called hemagglutinating virus ofJapan (HVJ). SeV is responsible for a highly transmissible
respiratory tract infection in mice, hamsters, guinea pigs,
rats, and occasionally pigs, with infection passing throughboth air and direct contact routes. The virus can be detectedin mouse colonies worldwide, generally in suckling toyoung adult mice. It is believed that the natural host of
Sendai virus is the mouse and that the virus is usually
nonpathogenic for humans. The SeV genome is organized
starting with the short 3’ leader region, followed by six
genes encoding the nucleocapsid protein (NP),phosphoprotein (P), matrix protein (M), fusion protein (F),hemagglutinin-neuraminidase protein (HN), and large
protein (L), and ending with a short 5’ trailer region. TheSeV now has been developed as gene transfer vectors forexpressing foreign genes to a wide range of mammaliancells and tissues with high efficiency.

Western blotting; 1:2,000
Immunoprecipitation; Not tested
Immunohistochemistry; Not tested
Immunocytochemistry; 1:500 (for methanol fixed cells)
Flow cytometry; Not tested**It is reported that this antibody can be used in Flowcytometry in the reference number 7).
Detailed procedure
PROTOCOLS

isprovidedin

thefollowing

The descriptions of the following protocols are examples.
Each user should determine the appropriate condition.
.

kDa
97
66

SOURCE: This antibody was whole serum. The rabbit

45

was immunized with wild type Sendai virus.

FORMULATION: 50 L volume of whole serum

31

containing 0.09% NaN3.
*Azide may react with copper or lead in plumbing system to form
explosive metal azides. Therefore, always flush plenty of water
when disposing materials containing azide into drain.

STORAGE: This antibody solution is stable for one year

Western blot analysis of Sendai virus
protein in recombinant GFP/dF-SeV
using PD029.

from the date of purchase when stored at 4°C.

REACTIVITY: This antibody reacts with Sendai virus

The descriptions of the following protocols are examples.
Each user should determine the appropriate condition.

on Western blotting and Immunocytochemistry.

PROTOCOLS:

SPECIES CROSS REACTIVITY:
Species
Recombinant

Human

Mouse

Rat

SDS-PAGE & Western Blotting
Other

Not tested Not tested Not tested GFP/dF-SeV

Reactivity onWB+INTENDED USE:
For Research Use Only. Not for use in diagnostic procedures.

1) Mix the sample with Laemmli’s sample buffer.
2) Boil the samples for 3 minutes and centrifuge. Load 20 Lof sample per lane on a 1-mm-thick SDS-polyacrylamide
gel and carry out electrophoresis.
3) Blot the protein to a polyvinylidene difluoride (PVDF)
membrane at 1 mA/cm2 for 1 hour in a semi-dry transfer
system (Transfer Buffer: 25 mM Tris, 190 mM glycine,
20% MeOH). See the manufacturer's manual for precisetransfer procedure.

MEDICAL & BIOLOGICAL LABORATORIES CO., LTD.
URL https://ruo.mbl.co.jp

e-mail support@mbl.co.jp

PD029
Lot 012~Page 24) To reduce nonspecific binding, soak the membrane in 5%skimmed milk (in PBS, pH 7.2) for 1 hour at room
temperature, or overnight at 4°C.
5) Incubate the membrane for 1 hour at room temperature
with primary antibody diluted with 1% skimmed milk (inPBS, pH 7.2) as suggested in the APPLICATIONS. (Theconcentration of antibody will depend on the conditions.)
6) Wash the membrane with PBS-T [0.05% Tween-20 inPBS] (5 minutes x 3 times).
7) Incubate the membrane with 1:10,000 Anti-IgG (Rabbit)
pAb-HRP (MBL; code no. 458) diluted with 1% skimmed
milk (in PBS, pH 7.2) for 1 hour at room temperature.
8) Wash the membrane with PBS-T (5 minutes x 3 times).
9) Wipe excess buffer off the membrane, and incubate
membrane with an appropriate chemiluminescencereagent for 1 minute.
10) Remove extra reagent from the membrane by dabbing
with a paper towel, and seal it in plastic wrap.
11) Expose the membrane onto an X-ray film in a dark roomfor 10 minutes. Develop the film under usual settings. The
conditions for exposure and development may vary.

10) Iwadate, Y., et al., Clin. Cancer Res. 11, 3821-3827 (2005)
11) Kato, M., et al., Vaccine 23, 3166-3173 (2005)12) Sasaki, K., et al., Gene Ther. 12, 203-210 (2005)
13) Inoue, M., et al., J. Gene Med. 6, 1069-1681 (2004)
14) Ferrari, S, et al., Gene Ther. 11, 1659-1664 (2004)
15) Matano, T., et al., J. Exp. Med. 199, 1709-1718 (2004)
16) Kinoh, H., et al., Gene Ther. 11, 1137-1145 (2004)17) Shirakura, M., et al., Gene Ther. 11, 784-790 (2004)
18) Okano, S., et al., Gene Ther. 10, 1381-1391 (2003)
19) Inoue, M., et al., J. Virol.77, 6419-6429 (2003)
20) Jin, C. H., et al., Gene Ther. 10, 272-277 (2003)
21) Tokusumi, T., et al., Virus Res. 86, 33-38 (2002)22) Masaki, I., et al., Circ. Res. 90, 966-973 (2002)
23) Yamashita, A., et al., J. Immunol. 168, 450-457 (2002)24) Shiotani, A., Gene Ther. 8, 1043-1050 (2001)25) Masaki, I., et al., FASEB J. 15, 1294-1296 (2001)
26) Yonemitsu, Y., et al., Nat. Biotechnol. 18, 970-973 (2000)
27) Li, H. O., et al., J. Virol. 74, 6564-6569 (2000)
28) Kato, A., et al., Genes Cells 1, 569-579 (1996)29) Inoue, M., and Hasegawa, M., Biotherapy 19, 85-92 (2005)(Positive control for Western blotting; Recombinant)Immunocytochemistry1) Wash the cells twice with pre-chilled PBS, and then fix
the cells by immersing the slide in methanol pre-cold at-20°C for 10 minutes on ice.
2) Allow the cells air dry, and then wash the cells twice withPBS for 3-5 minutes each.
3) Add the primary antibody diluted with PBS as suggested
in the APPLICATIONS onto the cells and incubate for
45 minutes at 37°C. (Optimization of antibody
concentration or incubation condition is recommended ifnecessary.)4) Wash as in step 2).
5) Add FITC-conjugated anti-rabbit IgG antibody diluted
with PBS onto the cells. Incubate for 30 minutes at room
temperature. Keep out light by aluminum foil.
6) Wash as in step 2).
7) Wipe excess liquid off the slide but take care not to touch
the cells. Never leave the cells to dry.
8) Promptly add mounting medium onto the slide, then put acover slip on it.
(Positive control for Immunocytochemistry; Transfectant)

REFERENCES:
1) Cheng, D., et al., J. Biol. Chem. 293, 5975-5986 (2018) [WB]
2) Yoshizumi, T., et al., Sci. Rep. 7, 5379 (2017) [WB]
3) Chan, Y. K., and Gack, M. U., Nat. Immunol. 17, 523-530(2016) [WB]
4) Zeltner, N. et al., Nat. Med. 22, 1421-1427 (2016) [IC]5) Choi, J. et al., Nat. Biochem. 33, 1173-1181 (2015) [IC]6) Lee, D. F., et al., Cell 161, 240-254 (2015) [IC]7) Hach, J. C., et al., J. Virol. 87, 9923-9927 (2013) [FCM]
8) Macarthur, C. C., et al., Stem Cells Int. 564612 (2012) [IC]9) Griesenbach, U., et al., Curr. Opin. Mol. Ther. 7, 346-52 (2005)
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