Anti-IZUMO1 antibody, rabbit serum
Product code73-042

Size

100 µlStorage

Store 4℃ for short term For long term storage store at -20℃.
Aliquot to avoid repeated freezing and thawing.

Concentration

N/A

Buffer

0.1% sodium azide

PurityRabbit antiserum

Immunogen

KLH-conjugated synthetic peptides corresponding to the following three regionsof human IZUMO 1.
[A] C+KSLEKDYLPGHLDA[B] C+TQVPKEKATDSRQQ[C] C+ATTESSISLQPLQ

IsotypeRabbit IgG

Reactivity

Human and mouse.

Not tested with other species.
Special notes

N/AApplication

1. Western blotting (1/1,000 dilution)
2. Immunofluorescence staining (1/100~1/300 dilution)
3. Immunohistochemistry (1/100 dilution)
4. Inhibition of sperm fusion with egg

Background

Essential sperm cell-surface protein required for fertilization by acting as aligand for FOLR4/JUNO receptor on egg. The IZUMO1:FOLR4/JUNO
interaction is a necessary adhesion event between sperm and egg that isrequired for fertilization but is not sufficient for cell fusion. The ligand-receptor
interaction probably does not act as a membrane 'fusogen'Molecular mass: 44,885 Da with 307 amino acids. Post-translationalmodification; Processing of N-terminal signal peptide with 21 amino acids. NGlucosylation and phosphorylation.
Expression: This gene has expression in 4 organs : EMAPA:18202: epidermis ,EMAPA:16105: heart , MA:0000412: seminiferous tubule , MA:0000411: testis

Data Link

UniProtKB Q8IYV9 (human IZUMO1),

UniProtKB Q9D9J7 (mouse IZUMO1)

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
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Data Images: Anti-IZUMO1 antibody, rabbit serum

Fig.1 Identification of IZUMO1 protein in the lysate of human sperm by western blotting with antiIZUMO1 antibody.
Proteins in the lysate (20 µg) was separated on SDS-PAGE, blotted to PVDF membrane and reactedwith anti-human IZUMO1 antibody at 1/1,000 dilution.

Fig.2 Analysis of IZUMO1 protein in the lysates of mouse sperm by western blotting with polyclonalanti- IZUMO1 antibody.
Proteins in the lysates (10 µg) was separated on SDS-PAGE (10~20% gradient gel), blotted to PVDF
membrane and reacted with the polyclonal anti-IZUMO1 antibody at 1/1,000 dilution.

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Fig.3 Immunostaining of IZUMO1 in human sperm using polyclonal anti-IZUMO1 antibody.
Human sperm on slide was incubated with polyclonal anti-IZUMO1 antibody at 1/100 dilution andreacted with a second antibody,

Alexa Fluor 594-conjugated anti-rabbit IgG antibody at 1/1,000(lower panel).

Fig.4. Immunohistochemistry of mouse testis using anti-IZUMO1 antibody.
Formalin-fixed and paraffin-embedded mouse testis
Deparaffinization by LemosolRA (#122-03991,Wako, Osaka)
Rehydration

100% EtOH, 95%, 90%, 70%, DW

Antigen retrieval

Histo/Zyme (Cat.# k046; Diagnostic BioSystems)

Washing

PBST (0.25% triton X-100/PBS-)

Blocking

10 % FBS / PBST 30 min

1st antibody

1/100 dilution in PBS- 4℃ O/NWashing

PBS- 5 min, 3 times

2nd antibody

1,000 dilution, 60 min (Alexa Flour-488 goat anti-rabbit IgG (H&L),#1166843 ; Molecular Probes)

Washing

PBS- 5 min, 3 times

DAPI

1.0μg/mL DAPI in TBS 10 min

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Reference: This antibody was described and used in the following publication.
1. Inoue N. et al. (2005) The immunoglobulin superfamily protein Izumo is required for sperm tofuse with eggs. Nature. 434:234-8. PubMeD 15759005

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