Anti-PDILT antibody, rabbit serum, KO-Validated
Product code73-051Size

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

ImmunogenTwo synthetic peptides corresponding to the C-terminal regions of mouse PDILT,C+IRKPEEPERRKETA (550-563) and C+QPKEQPKPERKLEV (571-584),
respectively, conjugated with KLH

IsotypeRabbit IgG

ReactivityMouse.
Not tested with other species.

Special notes

Validation: knock-out mouse

Application

1. Western blotting (1/1,000 dilution)
2. Immunoprecipitation (1/100)
3. Immunohistochemistry (1/100-1/500)

Background

PDILT cooperates with the testis-specific calreticulin-like chaperone, calsperin(CALR3), in the endoplasmic reticulum and plays an indispensable role in thedisulfide-bond formation and folding of ADAM3. Pdilt(-/-) mice were maleinfertile because ADAM3 could not be folded properly and transported to thesperm surface without the PDILT/CALR3 complex.
Molecular mass:67,759 with 588 amino acids. N-glucosylated

Data Link

UniProtKB Q9DAN1 Mouse PDILT,Gene ID 71830 Mouse PDILT

Please note: All products are FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC
PROCEDURES. NOT FOR MILITARY USE.
1 / 5

BioAcademia,Inc. Tel. 81-6-6877-2335

Fax. 81-6-6877-2336 info@bioacademia.co.jp https://www.bioacademia.co.jp/en/

Data Images: 73-051 Anti-PDILT antibody, rabbit serum, KO-Validated

Fig. 1. Validation of specificity of the anti-PDILT antibody with knockout mice testis extracts.
Samples wild-type, hybrid and knock-out were reacted with anti-PDILT antibody at 1/1,000 dilution.
α-Tubulin was used as a control.

Fig.2

Western blotting analysis of PDILT expression in various tissues with anti-PDILT antibody.

Proteins were extracted from various tissues with lysis buffer containing Triton X-100 and subjected
to western blot analysis. Tissue proteins (30 µg) and sperm protein (10 µg) were reacted with antiPDILT antibody at 1/1,000 dilution.

2 / 5

BioAcademia,Inc. Tel. 81-6-6877-2335

Fax. 81-6-6877-2336 info@bioacademia.co.jp https://www.bioacademia.co.jp/en/Fig.3 Western blot analysis of PDILT protein in the lysates of mouse testis and sperm with antiPDILT antibody.
Proteins in the lysates (10 μg) were separated on SDS-PAGE, electro-blotted to PVDF membrane andreacted with anti-PDILT antibody at 1/1,1000 dilution. Anti-rabbit IgG conjugated with HRP wasused at 10,000 dilution.

Fig.4. Immunoprecipitation of PDILT protein from mouse testis extracts. Extracts (100 μg) of wildtype (+/+) and Pdilt (-/-) mouse testes were immunoprecipitated with anti-PDILT antibody at 1/100
dilution and the precipitates were analyzed by western blotting using the same antibody at 1/1,000
dilution.
1. Input.
2. Wild-type.
3. Pdilt (-/-)

3 / 5

BioAcademia,Inc. Tel. 81-6-6877-2335

Fax. 81-6-6877-2336 info@bioacademia.co.jp https://www.bioacademia.co.jp/en/

Fig.5. Immunofluorescence staining of mouse testicular sections with anti-PDILT antibody.
Testes were collected from adult mice and were fixed in 4% paraformaldehyde/PBS, cryopreserved in
garaded 10-30% sucrose, and embedded in a TissueTek OCT compound (Sakura Finetechnical, Tokyo).
Frozen sections (5 μm) were mounted on APS-coated glass slides. After washing and blocking, theslides were reacted with anti-PDILT antibody at 1/100 dilution. As a secondary antibody, Alexa Fluor
546 conjugated anti-rabbit IgG antibody was used.
PDILT was detected only in spermatid.

Fig.6 Immunohistochemical analysis of paraffin embedded mouse testis tissue labelling PDILT with
PDILT antibody.
Deparaffinization LemosolRA (#122-03991,Wako, Osaka)
Rehydration
Antigen retrieval

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

Washing

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

Blocking

10 % FBS / PBST

1st antibody

1/1,000 dilution in PBS- 4℃ O/NWashingPBS-

2nd antibody

1,000 dilution, 60 min

washing PBS-

5 min X 3

DAPI

1.0μg/mL DAPI in TBS

30 min10 minWashingPBSMount ImmunoSelect Antifading Mounting Medium (SCR-38447; Dianova)
4 / 5

BioAcademia,Inc. Tel. 81-6-6877-2335

Fax. 81-6-6877-2336 info@bioacademia.co.jp https://www.bioacademia.co.jp/en/

Reference: This antibody was described and used in the following publication.
1. Tokuhiro K. et al (2012). Protein disulfide isomerase homolog PDILT is required for quality controlof sperm membrane protein ADAM3 and male fertility. Proc Natl Acad Sci U S A. 109:3850-5.
WB, IP. IHC. Free access.

5 / 5

BioAcademia,Inc. Tel. 81-6-6877-2335

Fax. 81-6-6877-2336 info@bioacademia.co.jp https://www.bioacademia.co.jp/en/