Anti-CD52 antibody, rabbit serum
Product code73-030

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

ImmunogenSynthetic peptide corresponding to 29-48 amino acids of mouse CD52, CAASGTNKNSTSTKKTPLKSG , conjugated with KLH

IsotypeRabbit IgG

Reactivity

Mouse.
Likely to react with rat due to the sequence identity.

Special notes

Validation: Specificity validated with KO mouse (Fig.2)

Application1. Western blotting (1/1,000 dilution)
2. Immunohistchemistry-P (1/100 dilution)
3. Immunofluorescence staining (1/100 dilution)

BackgroundCD52 may play a role in carrying and orienting carbohydrate, as well as having
a more specific role. Expressed on lymphohematopoietic tissues, including
thymus, spleen, and bone marrow, but not in liver, kidney, and brain.
Molecular mass: 7,798 Da with 74 amino acids. In mature form, propeptide isremoved, and GPI anchored and glycosylated.

Data Link

uniprot/Q64389 Mouse CD52

Gene ID 23833 Mouse CD52

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: 73-030 Anti-CD52 antibody, rabbit serum

Fig.1 Western blotting analysis of CD52 expression in various tissues with anti-CD52 antibody.
Testes, male reproductive ducts and sperm protein were extracted with lysis buffer containing Triton
X-100 and subjected to western blot analysis. Western blots containing equal amounts of tissue
proteins (30 µg) and sperm protein (10 µg) were reacted with anti-CD52 antibody at 1/1,000 dilution.

Fig.2. Western blot analysis of CD52 in cauda epididymal lysates and sperm lysates of wild type andCD52 deficient mice.

Cd52+/+ (+/+), Cd52+/– (+/–), and Cd52−/–(–/–).

Cauda epididymis and sperm from vas deferens were lysed in lysis buffer containing 1% TritonX-100.
Proteins (30 μg for cauda epididimis and 10 μg for sperm) were analyzed by western blotting with
anti-CD52 antibody at 1/1,000 dilution.

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Fig.3. Western blot analysis of CD52 in lysates of mouse
testis and sperm with anti-CD52 antibody.
Proteins in the lysates (10 μg) are separated on SDS-PAGE (10-20% gradient) and electro-blotted toPVDF membrane. The membrane was reacted with anti-CD52 antibody at 1/1,000 dilution. As thesecond antibody, anti-rabbit IgG antibody conjugated with HRP (ab97051) was used at 1/10,000
dilution. The numbers on the right are positions of protein size markers shown in kDa.

Fig.4. Immunohistochemistry of mouse testis using anti-CD52 antibody.
Formalin-fixed and paraffin-embedded mouse testis
Deparaffinization

LemosolRA (#122-03991,Wako, Osaka)

Rehydration

100% Et-OH, 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/NWashingPBS-

2nd antibody

1,000 dilution, 60 min (Alexa Flour-488 goat anti-rabbit IgG (H&L)

Washing PBS-5 min, 3 times

DAPI

1.0μg/mL DAPI in TBS

MountImmunoSelect Antifading Mounting Medium (SCR-38447; Dianova)

10 min3 / 4

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Fig. 5 Immunofluorescence staining of CD52 in NIH3T3 cells with anti-CD52 antibody.
The cells were fixed in 4% paraaformaldehyde over night
Permeabilization in 0.25% Triton X-100/PBS for 10 min
Blocking in 1.5% BSA/PBS for 30 min
1st antibodies diluted 1/100 by blocking buffer and incubated over night
2nd antibody,goat, anti-mouse IgG conjugated withAlex 488 (1/1000 dilution).
Nuclei were stained with DAPI
Reference: This antibody was described in Ref.1 and used in the following publications.
1. Yamaguchi R et al. (2008) Cd52, known as a major maturation-associated sperm membrane
antigen secreted from the epididymis, is not required for fertilization in the mouse. Genes
Cells. 13:851-61. PMID: 18782223 WB (mouse).

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