Anti-PAX6 antibody [EPR15858] is a rabbit recombinant monoclonal antibody that is used to detect PAX6 in ICC/IF, IHC-P, Western blot. Suitable for Human, Mouse, Rat samples.
- Validated on the Leica BOND™ RX automated IHC staining platform for PAX6 IHC
- Cited in over 50 publications
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
WB | ICC/IF | IHC-P | |
---|---|---|---|
Human | Tested | Tested | Tested |
Mouse | Tested | Tested | Expected |
Rat | Expected | Expected | Tested |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/1000 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/350 | Notes - |
Species Human | Dilution info 1/350 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1/500 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/500 | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
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PAX6 is a transcription factor with important roles in the development of the eye, nose, central nervous system, and pancreas. It competes with PAX4 for binding to a common element in the promoters of glucagon, insulin, and somatostatin. This supplementary information is collated from multiple sources and compiled automatically.
AN2, PAX6, Paired box protein Pax-6, Aniridia type II protein, Oculorhombin
Anti-PAX6 antibody [EPR15858] is a rabbit recombinant monoclonal antibody that is used to detect PAX6 in ICC/IF, IHC-P, Western blot. Suitable for Human, Mouse, Rat samples.
- Validated on the Leica BOND™ RX automated IHC staining platform for PAX6 IHC
- Cited in over 50 publications
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
IHC image of Pax6 staining in a formalin fixed, paraffin embedded normal human cerebellum tissue section, 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 20 mins. The section was then incubated with ab195045, 1/1000 dilution, 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.
Immunofluorescence analysis of 4% paraformaldehyde-fixed, 0.1% tritonX-100 Neuro-2a (mouse neuroblastoma) cells labeling PAX6 with ab195045 at 1/350, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/400 (green).
Confocal image showing cytoplasmic and nuclear staining on Neuro-2a cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/500 and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 (red).
The negative controls are as follows:-
-ve control 1: ab195045 at 1/350 followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/500 followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400.
Immunohistochemical analysis of paraffin-embedded Human retina tissue labeling PAX6 with ab195045 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Nuclear staining on human retina 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) (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.
Blocking/Dilution buffer: 5% NFDM/TBST.
The Transfected lysate spans the immunogenic region: aa1-aa422(47kDa).
All lanes: Western blot - Anti-PAX6 antibody [EPR15858] (ab195045) at 1/10000 dilution
Lane 1: PAX6 transfected 293T cell lysate at 10 µg
Lane 2: Non-transfected 293T cell lysate at 10 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 47 kDa
Observed band size: 32 kDa, 33 kDa, 47 kDa
Immunofluorescence analysis of 4% paraformaldehyde-fixed, 0.1% tritonX-100 Y79 (Human retinoblastoma cell line) cells labeling PAX6 with ab195045 at 1/350, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/400 (green).
Confocal image showing cytoplasmic and nuclear staining on Y79 cell line.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (anti-Tubulin mouse mAb) at 1/500 and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 (red).
The negative controls are as follows:-
-ve control 1: ab195045 at 1/350 followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500.
-ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 at 1/500 followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/400.
Blocking/Dilution buffer: 5% NFDM/TBST.
The 47kDa band represents the full length PAX6, we hypothesis the 32kDa & 33kDa bands represent the PAX6p32 & PAX6p33 fragments.
All lanes: Western blot - Anti-PAX6 antibody [EPR15858] (ab195045) at 1/1000 dilution
All lanes: Mouse eyeball lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 47 kDa
Observed band size: 32 kDa, 33 kDa, 47 kDa
Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling PAX6 with ab195045 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Nuclear staining on human pancreas 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) (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.
Blocking/Dilution buffer: 5% NFDM/TBST.
The 47kDa band represents the full length PAX6, we hypothesis the 32kDa & 33kDa bands represent the PAX6p32 & PAX6p33 fragments.
All lanes: Western blot - Anti-PAX6 antibody [EPR15858] (ab195045) at 1/10000 dilution
Lane 1: Y79 cell lysate at 20 µg
Lane 2: HeLa cell lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 47 kDa
Observed band size: 32 kDa, 33 kDa, 47 kDa
Immunohistochemical analysis of paraffin-embedded Human retinoblastoma tissue labeling PAX6 with ab195045 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) secondary antibody (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Nuclear staining on human retinoblastoma 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) (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 image of Pax6 staining in a formalin fixed, paraffin embedded normal rat cerebellum tissue section, 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 20 mins. The section was then incubated with ab195045, 1/1000 dilution, 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.
Western blot: Anti-PAX6 antibody [EPR15858] (ab195045) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab195045 was shown to bind specifically to PAX6. A band was observed at 50 kDa in wild-type A549 cell lysates with no signal observed at this size in PAX6 knockout cell line. To generate this image, wild-type and PAX6 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-PAX6 antibody [EPR15858] (ab195045) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: PAX6 knockout A549 cell lysate at 20 µg
Lane 3: HEK-293 cell lysate at 20 µg
Lane 4: MCF7 cell lysate at 20 µg
All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 50 kDa
ab195045 staining Pax6 in primary mouse neurons/glia, DIV1 (top row) and DIV14 (bottom row) both prepared from E18 mouse hippocampal brain area (obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. C57EHP). 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 ab195045 at 0.2 µg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, mouse monoclonal [DM1A] to alpha Tubulin, at 1/1000 dilution. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
A subset of cells showed staining in the nucleus at DIV1 (likely neuroprogenitor cells), while differentiated neurons (DIV14) where Pax6 negative.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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