Anti-Otx2 antibody

• IHC-P

Collaborator31270****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (AB114138)

IHC-P image of Otx2 staining on mouse E14 embryo using ab114138 (1 : 1000). The sections were deparffinized and subjected to heat mediated antigen retrieval using citric acid. The section were then blocked using 1% BSA for 10 mins at 21°C. ab114138 was diluted 1 : 1000 using TBS/BSA/Azide and incubated for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to Biotin (1 : 250).

This image is courtesy of Carl Hobbs (Kings College London, United Kingdom)

• IHC-P

Collaborator

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (AB114138)

IHC-P image of Otx2 antibody (ab114138) on paraffin-embedded mouse embryo sections. The image on the left is of a WT embryo, whilst on the right is an embryo where Otx2 has been replaced with human Otx1 (hOtx1^Otx2/Otx2). The sections underwent heat mediated antigen retrieval using Citrate Buffer, primary antibody was used at 1 : 500 dilution and Biocare Mach4 Kit was used to detect the primary antibody.

This image is courtesy of Prof. Antonio Simeone, Institute of Genetics and Biophysics, Naples, Italy.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (AB114138)

IHC image of Otx2 staining in a section of formalin-fixed paraffin-embedded normal mouse e14 foetal brain performed on a Leica BOND™ 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 20mins. The section was then incubated with ab114138, 2.5ug/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. The inset secondary-only control image is taken from an identical assay without primary antibody.

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.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (AB114138)

IHC image of Otx2 staining in a section of formalin-fixed paraffin-embedded normal mouse e14 foetal brain performed on a Leica BOND™ 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 20mins. The section was then incubated with ab114138, 2.5ug/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. The inset secondary-only control image is taken from an identical assay without primary antibody.

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.

• IHC-P

Collaborator31270****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Otx2 antibody (AB114138)

IHC-P image of Otx2 staining on Rat E12 embryo using ab114138 (1 : 1500). The sections were deparffinized and subjected to heat mediated antigen retrieval using citric acid. The section were then blocked using 1% BSA for 10 mins at 21°C. ab114138 was diluted 1 : 1500 using TBS/BSA/Azide and incubated for 2 hours at 21°C. The secondary antibody used was goat polyclonal to rabbit IgG conjugated to Biotin (1 : 250).

This image is courtesy of Carl Hobbs (Kings College London, United Kingdom)

• WB

Unknown

Western blot - Anti-Otx2 antibody (AB114138)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab114138 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-Otx2 antibody (ab114138) at 1 µg/mL

Lane 1:

E10 Mouse Embryo Brain Tissue Lysate at 10 µg

Lane 2:

E14 Mouse Embryo Brain Tissue Lysate at 10 µg

Lane 3:

E16 Ms Embryo Brain Tissue Lysate at 10 µg

Lane 4:

Western blot - Mouse eye tissue lysate - total protein (<a href='/en-us/products/tissue-lysates/mouse-eye-tissue-lysate-total-protein-ab4029'>ab4029</a>) at 10 µg

Lane 5:

Y79 (Human retinoblastoma cell line) Whole Cell Lysate at 10 µg

Lane 6:

WERI (Human Retinoblastoma) Whole Cell Lysate at 10 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 1/10000 dilution

Predicted band size: 32 kDa

Observed band size: 37 kDa,62 kDa

true

Exposure time: 12min

• WB

Unknown

Western blot - Anti-Otx2 antibody (AB114138)

Lane 1 represents the Otx1 Recombinat protein which was used as a negative control for ab114138. This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab114138 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-Otx2 antibody (ab114138) at 1 µg/mL

Lane 1:

OTX1 (Human) - Recombinant Protein at 0.1 µg

Lane 2:

OTX2 Transient Overexpression Lysate at 0.1 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 1/10000 dilution

Predicted band size: 32 kDa

Observed band size: 37 kDa,62 kDa

true

Exposure time: 8min