Localization of MAFB in adult mouse testis.

Double immunostaining of MAFB with SCP3 (ab97672 at a 1/400 dilution, O/N +4°C). Nuclei were counterstained with DAPI. The color of each marker is indicated above the images. All seminiferous tubules shown represent stage VII. US; undifferentiated spermatogonia. DS; differentiated spermatogonia. P; pachytene spermatocytes. Sp; spermatids. S; Sertoli cells. L; Leydig cells. MAFB was specifically detected in Leydig cells, Sertoli cells, and pachytene spermatocytes.

 

IHC image of SCP3 staining in a section of formalin-fixed paraffin-embedded mouse normal testis, using Mouse on Mouse detection kit, ab127055.

The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins, and incubated overnight at +4°C with ab97672 at 1 µg/ml dilution. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10mins at room temperature. The section was counterstained with hematoxylin and mounted with DPX.

The inset negative control image is taken from an identical assay without primary antibody.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

All lanes : Anti-SCP3 antibody [Cor 10G11/7] (ab97672) at 1 µg/ml

Lane 1 : Testis (Mouse) Tissue Lysate
Lane 2 : Human testis tissue lysate - total protein (ab30257)

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 28 kDa
Additional bands at: 125 kDa (possible non-specific binding)


Exposure time: 30 seconds

IHC-P image of SCP3 staining with ab97672 on tissue sections from adult mouse testis.

The sections were subjected to heat-mediated antigen retrieval using Dako antigen retrieval solution. The sections were then blocked with 5% milk for 30 minutes at 25°C, before incubation with ab97672 (1/200 dilution) for 16 hours at 4°C. The secondary was an Alexa-Fluor^®488 conjugated goat anti-mouse polyclonal, used at a 1/500 dilution.

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ab97672 staining SCP3 in mouse adult testis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).

Tissue was fixed with 4% paraformaldehyde and blocked with 5% BSA for 60 minutes at 25°C; antigen retrieval was by heat mediation in a 0.01M citrate buffer. Samples were incubated with primary antibody (1/200 in 1% BSA) for 1 hour at 22°C. A Biotin-conjugated Goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

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