IHC image of DAZL staining in mouse testis formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab34139, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

ab34139 at 1/100 staining human blastocyst cells by ICC/IF. The cells were paraformaldehyde fixed and blocked with 5% serum prior to incubation with the antibody for 18 hours. An Alexa-Fluor ® 488 conjugated secondary antibody was used.

See Abreview

The image, submitted as part of an Abreview, shows DAZL staining (green) of a human blastocyst. The antibody stains the cytoplasm of cells of the inner cell mass and trophoblast. DAZL is known to be located to the cytoplasm and DAZL transcripts have previously been discovered in the inner cell mass and trophblast (see PMID: 15879466).

The image shows staining of human testis tissue. The cytoplasm of Leydig cells and other cells in the seminiferous tubules stained positive. The antigen retrieval techniques used were Citrate at pH6 or Tris EDTA at pH9.

All lanes : Anti-DAZL antibody (ab34139) at 1 µg/ml

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

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size: 33 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?
Additional bands at: 130 kDa, 70 kDa, 95 kDa. We are unsure as to the identity of these extra bands.


Exposure time: 2 minutes

Ab34139 staining DAZL on juvenile marmoset testes by immunohistochemistry. The tissue was paraformaldehyde fixed and blocked in 1% milk for 1 hour at 25°C. The primary antibody was diluted 1/100 and incubated with the sample for 9 hours at 4°C in 5% milk. An Alexa Fluor 555 conjugated goat anti-rabbit antibody was used as the secondary. Nuclear staining shown in blue with DAPI.

See Abreview

Anti-DAZL antibody (ab34139) at 1 µg/ml + GST tagged Recombinant Human DAZL protein (ab112279) at 0.1 µg

Secondary
Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 33 kDa


Exposure time: 10 seconds