Anti-Sp8 antibody [EPR26224-86] - BSA and Azide free

• IHC

Supplier Data

Immunohistochemistry - Anti-Sp8 antibody [EPR26224-86] - BSA and Azide free (AB302916)

This data was developed using ab302915, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded A HEK-293T cells tr tissue labeling Sp8 with ab302915 at 1/5000 (0.106 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Nuclear staining on HEK-293T cells transfected with a human SP8 expression vector containing a myc-His tag cell pallet (image A). No staining on HEK-293T cells transfected with empty vector containing a his tag cell pallet (image B). The section was incubated with ab302915 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC

Supplier Data

Immunohistochemistry - Anti-Sp8 antibody [EPR26224-86] - BSA and Azide free (AB302916)

This data was developed using ab302915, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human prostate tissue labeling Sp8 with ab302915 at 1/50 (10.6 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Nuclear staining on human prostate. The section was incubated with ab302915 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC

Supplier Data

Immunohistochemistry - Anti-Sp8 antibody [EPR26224-86] - BSA and Azide free (AB302916)

This data was developed using ab302915, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Rat brain tissue labeling Sp8 with ab302915 at 1/200 (2.65 ug/ml) followed by a LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. (A) Nuclear staining on olfactory bulb of rat brain at low-powered (magnification, x10) and (B) nuclear staining on rat olfactory bulb at high-powered (magnification, x400). The section was incubated with ab302915 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is LeicaDS9800 (Bond™ Polymer Refine Detection) was used at Ready to use dilution. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-Sp8 antibody [EPR26224-86] - BSA and Azide free (AB302916)

This data was developed using ab302915, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat olfactory bulb (fresh) tissue labeling Sp8 with ab302915 at 1/500 (1.06 ug/ml) dilution followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Positive staining on rat olfactory bulb. The nuclear counterstain was DAPI (Blue). The section was incubated with ab302915 for 60 mins at room temperature. The section was then mounted using Fluoromount®.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution.<\p> .

• WB

Supplier Data

Western blot - Anti-Sp8 antibody [EPR26224-86] - BSA and Azide free (AB302916)

This data was developed using ab302915, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST

Low expression :  spleen, lung (consistent with the predicted expression pattern).

Bands below 60 kDa may due to degradation.

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-Sp8 antibody [EPR26224-86] (<a href='/en-us/products/primary-antibodies/sp8-antibody-epr26224-86-ab302915'>ab302915</a>) at 1/1000 dilution

Lane 1:

Rat olfactory bulb tissue lysate at 20 µg

Lane 2:

Rat spleen tissue lysate at 20 µg

Lane 3:

Rat lung tissue 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 1/50000 dilution

Predicted band size: 48 kDa

Observed band size: 30-60 kDa

true

Exposure time: 180s