Anti-PROX1 antibody [EPR19273] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PROX1 antibody [EPR19273] - BSA and Azide free (AB236026)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma cell line from bone marrow) and HEK-293 (Human epithelial cell line from embryonic kidney) cells labeling PROX1 with ab199359 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing nuclear staining on SH-SY5Y cell line.

Negative expression in HEK-293 cells. HEK-293 doesn't express PROX1. PMID : 18948102.

The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG (AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab199359 at 1/500 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab7291  at 1/1000 dilution followed by ab150077  at 1/1000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab199359).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PROX1 antibody [EPR19273] - BSA and Azide free (AB236026)

Immunohistochemical analysis of paraffin-embedded rat hippocampus tissue labeling PROX1 with ab199359 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus staining on neurons of rat hippocampus is observed.

Counter stained with Hematoxylin.

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

This data was using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab199359).

Heat mediated antigen retrieval was performed 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-PROX1 antibody [EPR19273] - BSA and Azide free (AB236026)

Immunohistochemical analysis of paraffin-embedded mouse hippocampus tissue labeling PROX1 with ab199359 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Nucleus staining on neurons of mouse hippocampus is observed.

Counter stained with Hematoxylin.

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

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab199359).

Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ChIC/CUT&RUN-seq

Supplier Data

ChIC/CUT&RUN sequencing - Anti-PROX1 antibody [EPR19273] - BSA and Azide free (AB236026)

ChIC/CUT&RUN was performed using a pAG-MNAse at a final concentration of 700 ng/mL, 2.5 x 10^5 HepG2 (Human liver hepatocellular carcinoma cell line) cells and 5 µg of ab199359 [EPR19273]. The resulting DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 10 million reads. The negative IgG control ab172730 is also shown.

Additional screenshots of mapped reads can be found in the Protocol booklet in the Product Protocol section.

The University of Geneva owns patents relevant to ChIC (Chromatin Immuno-Cleavage) methods.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab199359).