Anti-DAZL antibody [EPR21028] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DAZL antibody [EPR21028] - BSA and Azide free (AB228135)

Immunohistochemical analysis of paraffin-embedded human testis tissue labeling DAZL with ab215718 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in primary spermatocytes of human testis (PMID : 24746554) is observed. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215718).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-DAZL antibody [EPR21028] - BSA and Azide free (AB228135)

This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab215718). ab215718 staining DAZL in mES cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab215718 at 1µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue). Also suitable in cells fixed with 100% methanol (5 min). Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-DAZL antibody [EPR21028] - BSA and Azide free (AB228135)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton-X-100 permeabilized frozen mouse testis tissue labeling DAZL with ab215718 at 1/300 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasmic staining in the pachytene spermatocytes and secondary spermatocytes, not present in Sertoli, Leydig or myoid cells (PMID : 24746554).

The nuclear counter stain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215718).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-DAZL antibody [EPR21028] - BSA and Azide free (AB228135)

Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton-X-100 permeabilized frozen rat testis tissue labeling DAZL with ab215718 at 1/60 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Cytoplasmic staining in the pachytene spermatocytes and secondary spermatocytes, not present in Sertoli, Leydig or myoid cells (PMID : 24746554).

The nuclear counter stain is DAPI (blue).

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215718).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DAZL antibody [EPR21028] - BSA and Azide free (AB228135)

Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling DAZL with ab215718 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in spermatogonia and primary spermatocytes of mouse testis (PMID : 24746554) is observed. Counterstained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215718).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DAZL antibody [EPR21028] - BSA and Azide free (AB228135)

Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling DAZL with ab215718 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining in spermatogonia and primary spermatocytes of rat testis (PMID : 24746554) is observed. Counterstained with hematoxylin

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215718).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IP

Supplier Data

Immunoprecipitation - Anti-DAZL antibody [EPR21028] - BSA and Azide free (AB228135)

DAZL was immunoprecipitated from 0.35 mg mouse testis lysate with ab215718 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab215718 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

Lane 1 : Mouse testis lysate 10 μg (Input).

Lane 2 : ab215718 IP in mouse testis lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab215718 in mouse testis lysate.

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 3 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215718).

All lanes:

Immunoprecipitation - Anti-DAZL antibody [EPR21028] (<a href='/en-us/products/primary-antibodies/dazl-antibody-epr21028-ab215718'>ab215718</a>)

Predicted band size: 33 kDa

false

• WB

Unknown

Western blot - Anti-DAZL antibody [EPR21028] - BSA and Azide free (AB228135)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215718).

Blocking/Dilution buffer : 5% NFDM/TBST.

The blot was developed on a BIO-RAD^® ChemiDoc™ MP instrument.

All lanes:

Western blot - Anti-DAZL antibody [EPR21028] - BSA and Azide free (ab228135) at 1/1000 dilution

Lane 1:

Mouse testis lysate at 20 µg

Lane 2:

Rat testis lysate at 20 µg

Lane 3:

Human testis lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 33 kDa

Observed band size: 38 kDa

true

Exposure time: 3min

• WB

AbReview66346****

Western blot - Anti-DAZL antibody [EPR21028] - BSA and Azide free (AB228135)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab215718).

10% gel.

Blocked with 5% milk for 1 hour at room temperature.

Incubated with the primary antibody for 12 hours at 4°C in TBST.

All lanes:

Western blot - Anti-DAZL antibody [EPR21028] (<a href='/en-us/products/primary-antibodies/dazl-antibody-epr21028-ab215718'>ab215718</a>) at 1/1000 dilution

Lane 1:

Mouse testis tissue lysate at 25 µg

Lane 2:

DAZL knockout mouse testis tissue lysate at 25 µg

Secondary

All lanes:

HRP-conjugated horse anti-rabbit IgG at 1/10000 dilution

Predicted band size: 33 kDa

Observed band size: 37 kDa

true

Exposure time: 3s

This image is courtesy of an anonymous collaborator abreview.