PAX6 Recombinant Rabbit Monoclonal Antibody [SD081-03]
cat.: ET1612-58
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, FC, mIHC, IF-Cell, IF-Tissue
Clonality: Monoclonal
Clone number: SD081-03
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 47 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human PAX6 aa 373-422 / 422.
Positive control: HeLa cell lysate, 293T cell lysate, RAW264.7 cell lysate, Mouse cerebellum tissue lysate, Rat cerebellum tissue lysate, human pancreas tissue, mouse cerebellum tissue, mouse pancreas tissue, rat pancreas tissue, mouse eyeball tissue, rat eyeball tissue, rat cerebellum tissue, RAW264.7.
Subcellular location: Nucleus.
Recommended Dilutions:
  WB
  FC
  IHC-P
  mIHC
  IF-Cell
  IF-Tissue
1:5,000
1:1,000
1:200-1:2,000
1:1,000
1:100
1:800
Uniprot #: SwissProt: P26367 Human | P63015 Mouse | P63016 Rat
Alternative names: AN 2 AN AN2 Aniridia type II protein D11S812E FVH1 KIAA0552 Leucine zipper putative tumor suppressor 3 LZTS3 MGC17209 MGDA Oculorhombin Paired box 6 Paired box gene 6 (aniridia keratitis) Paired Box Gene 6 Paired box homeotic gene 6 Paired box protein Pax-6 Paired box protein Pax6 PAX 6 PAX6 PAX6_HUMAN ProSAP-interacting protein 1 PROSAPIP1 Sey WAGR
Images
ET1612-58_1.jpg Fig1: Western blot analysis of PAX6 on different lysates with Rabbit anti-PAX6 antibody (ET1612-58) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: MCF7 cell lysate (negative)
Lane 3: 293T cell lysate
Lane 4: RAW264.7 cell lysate
Lane 5: Mouse cerebellum tissue lysate
Lane 6: Rat cerebellum tissue lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 47 kDa
Observed band size: 47 kDa

Exposure time: Lane 1-3: 1 minute; Lane 4-6: 3 minutes; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1612-58) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1612-58_2.jpg Fig2: Fluorescence multiplex immunohistochemical analysis of mouse brain (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-NeuN (ET1602-12, red), anti-PAX6 (ET1612-58, green), anti-CD34 (ET1606-11, gray), anti-MAP2 (HA500177, magenta) and anti-TBR1 (ET1702-97, yellow) on mouse brain. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in five rounds of staining: in the order of ET1602-12 (1/5,000 dilution), ET1612-58 (1/1,000 dilution), ET1606-11 (1/2,000 dilution), HA500177 (1/5,000 dilution) and ET1702-97 (1/1,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Olympus VS200 Slide Scanner.
ET1612-58_3.jpg Fig3: Fluorescence multiplex immunohistochemical analysis of mouse pancreas (Formalin/PFA-fixed paraffin-embedded sections). Panel A: the merged image of anti-Cytokeratin 19 (ET1601-6, White) and anti-PAX6 (ET1612-58, Violet) on pancreas. HRP Conjugated UltraPolymer Goat Polyclonal Antibody HA1119/HA1120 was used as a secondary antibody. The immunostaining was performed with the Sequential Immuno-staining Kit (IRISKit™MH010101, www.luminiris.cn). The section was incubated in two rounds of staining: in the order of ET1601-6 (1/5,000 dilution) and ET1612-58 (1/1,000 dilution) for 20 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 30 mins at 95℃. DAPI (blue) was used as a nuclear counter stain. Image acquisition was performed with Zeiss Observer 7 Inverted Fluorescence Microscope.
ET1612-58_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human pancreas tissue with Rabbit anti-PAX6 antibody (ET1612-58) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-58) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1612-58_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue with Rabbit anti-PAX6 antibody (ET1612-58) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-58) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1612-58_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse pancreas tissue with Rabbit anti-PAX6 antibody (ET1612-58) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-58) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1612-58_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat pancreas tissue with Rabbit anti-PAX6 antibody (ET1612-58) at 1/2,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-58) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1612-58_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse eyeball tissue with Rabbit anti-PAX6 antibody (ET1612-58) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-58) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1612-58_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat eyeball tissue using anti-PAX6 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-58, 1/200) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1612-58_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue with Rabbit anti-PAX6 antibody (ET1612-58) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1612-58) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1612-58_11.jpg Fig11: Immunocytochemistry analysis of RAW264.7 cells labeling PAX6 with Rabbit anti-PAX6 antibody (ET1612-58) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-PAX6 antibody (ET1612-58) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1612-58_12.jpg Fig12: Immunofluorescence analysis of paraffin-embedded mouse cerebellum tissue labeling PAX6 with Rabbit anti-PAX6 antibody (ET1612-58) at 1/800 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (ET1612-58, green) at 1/800 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).
ET1612-58_13.jpg Fig13: Flow cytometric analysis of RAW264.7 cells labeling PAX6.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1612-58, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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