Name: Anti-PAX6 antibody [EPR15858]
SKU: ab195045
Rating: 5 (9 reviews)

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

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (AB195045), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	ICC/IF	IHC-P
Human	Tested	Tested	Tested
Mouse	Tested	Tested	Expected
Rat	Expected	Expected	Tested

Tested
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.

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/350	Notes -
Species Human	Dilution info 1/350	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

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.

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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12 products for Alternative Product

Target data

See full target information PAX6

Function

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.

Alternative names

AN2, PAX6, Paired box protein Pax-6, Aniridia type II protein, Oculorhombin

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR15858
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Product Specifications
Anti-PAX6 antibody [EPR15858] (ab195045) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in ICC/IF, IHC-P, WB in human, mouse, rat samples.
Anti-PAX6 antibody [EPR15858] (ab195045) specifically detects PAX6 (UniProt ID: P26367; Molecular weight: 47kDa) and is sold in 100 µL and 1 mL selling sizes.

Quality and Validation
Abcam's high quality manufacturing and validation processes ensure Anti-PAX6 antibody [EPR15858] (ab195045) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.
Anti-PAX6 antibody [EPR15858] (ab195045) has been cited over 57 times in peer reviewed journals and is trusted by the scientific community.
Anti-PAX6 antibody [EPR15858] (ab195045) has 8 independent reviews from customers.

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:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

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.

12 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)
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.
Immunocytochemistry/ Immunofluorescence - Anti-PAX6 antibody [EPR15858] (ab195045)
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)
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.
Western blot - Anti-PAX6 antibody [EPR15858] (ab195045)
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
Secondary
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
Immunocytochemistry/ Immunofluorescence - Anti-PAX6 antibody [EPR15858] (ab195045)
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.
Western blot - Anti-PAX6 antibody [EPR15858] (ab195045)
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
Secondary
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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)
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.
Western blot - Anti-PAX6 antibody [EPR15858] (ab195045)
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
Secondary
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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PAX6 antibody [EPR15858] (ab195045)
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)
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
Secondary
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
Immunocytochemistry/ Immunofluorescence - Anti-PAX6 antibody [EPR15858] (ab195045)
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).