Anti-Nkx3.1 antibody [EPR16653] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nkx3.1 antibody [EPR16653] - BSA and Azide free (AB251217)

This data was developed using ab196020, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human prostate hyperplasia tissue labeling Nkx3.1 with ab196020 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Nuclear staining on Human prostate hyperplasia tissue is observed. Counter stained with Hematoxylin.

Negative control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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-Nkx3.1 antibody [EPR16653] - BSA and Azide free (AB251217)

This data was developed using ab196020, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling Nkx3.1 with ab196020 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution. Human testis tissue acting as a negative control for Nkx3.1. Counter stained with Hematoxylin.

Negative control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

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

• mIHC

Lab

Multiplex immunohistochemistry - Anti-Nkx3.1 antibody [EPR16653] - BSA and Azide free (AB251217)

Fluorescence multiplex immunohistochemical analysis of human prostate gland tissue (formalin/PFA-fixed paraffin-embedded section). Panel A : merged staining of anti-Prostate Specific Antigen (ab76113, red; Opal™690), anti-Cytokeratin 5 (ab236216, green; Opal™520) and anti-Nkx3.1 (ab196020, gray; Opal™570) on human prostate gland tissue. Panel B : anti-Prostate Specific Antigen stained on cytoplasm of luminal cells. Panel C : anti-Cytokeratin 5 stained on basal cells. Panel D : anti-p63 stained on nucleus of luminal cells. Opal Polymer HRP Ms + Rb was used as a secondary antibody. The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. The section was incubated in three rounds of staining : in the order of ab76113 (1/2000), ab236216 (1/400) and ab196020 (1/2000) for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) was used for 20 mins. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Leica SP8 confocal microscope. This data was developed using ab196020, the same antibody clone in a different buffer formulation.

• WB

Supplier Data

Western blot - Anti-Nkx3.1 antibody [EPR16653] - BSA and Azide free (AB251217)

This data was developed using ab196020, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Lane 2, PC-3 (Human prostate cancer cell line) is acting as a negative control for Nkx3.1. The 32kDa band may represent the glycosylated form.

All lanes:

Western blot - Anti-Nkx3.1 antibody [EPR16653] (<a href='/en-us/products/primary-antibodies/nkx31-antibody-epr16653-ab196020'>ab196020</a>) at 1/1000 dilution

Lane 1:

LNCaP (Human prostate cancer cell line) cell lysate at 10 µg

Lane 2:

PC-3 (Human prostate cancer cell line) cell lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 26 kDa

Observed band size: 32 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Nkx3.1 antibody [EPR16653] - BSA and Azide free (AB251217)

This data was developed using ab196020, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Nkx3.1 antibody [EPR16653] (<a href='/en-us/products/primary-antibodies/nkx31-antibody-epr16653-ab196020'>ab196020</a>) at 1/1000 dilution

All lanes:

Human prostate lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 26 kDa

Observed band size: 32 kDa

false

Exposure time: 3min