Anti-DGKZ/DGK-zeta antibody [EPR22040-80] - BSA and Azide free

• mIHC

Lab

Multiplex immunohistochemistry - Anti-DGKZ/DGK-zeta antibody [EPR22040-80] - BSA and Azide free (AB239790)

This data was developed using ab239081, the same antibody clone in a different buffer formulation.

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human cerebellum staining of Calbindin with ab108404 at a 1/1000 dilution, DGKZ/DGK-zeta with ab239081 at 1/5000 dilution, and GABA A Receptor alpha 6 with ab300069 at a 1/500 dilution, followed by a ready to use secondary antibody Opal Polymer HRP Ms + Rb.

Panel A : merged staining of anti-GABA A Receptor alpha 6 (gray; Opal™690), anti-DGKZ/DGK-zeta (green; Opal™520) and anti-Calbindin (magenta; Opal™570) on human cerebellum.
Panel B : anti-Calbindin stained on all layers of cerebellum.
Panel C : anti-DGKZ/DGK-zeta stained on Purkinje cells.
Panel D : anti-GABA A Receptor alpha 6 stained on the granular layer.
Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab300069, ab239081, and ab108404 for 30 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, epitope retrieval solution2) for 20 mins.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DGKZ/DGK-zeta antibody [EPR22040-80] - BSA and Azide free (AB239790)

Immunohistochemical analysis of paraffin-embedded human cerebellum tissue labeling DGKZ/DGK-zeta with ab239081 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining on Purkinje cells of human cerebellum (PMID : 14511325; PMID : 24119575) 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) Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab239081).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-DGKZ/DGK-zeta antibody [EPR22040-80] - BSA and Azide free (AB239790)

Immunofluorescent analysis of 100% methanol-fixed Jurkat (human T cell leukemia cell line from peripheral blood) cells labeling DGKZ/DGK-zeta with ab239081 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining in Jurkat cell line.

Methanol is recommended for fixation as weak signal was observed using 4% PFA.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody 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 (ab239081).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-DGKZ/DGK-zeta antibody [EPR22040-80] - BSA and Azide free (AB239790)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized Jurkat (human T cell leukemia cell line from peripheral blood) cell line labeling DGKZ / DGK-zeta with ab239081 at 1/600 (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab239081).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-DGKZ/DGK-zeta antibody [EPR22040-80] - BSA and Azide free (AB239790)

Immunofluorescent analysis of 100% methanol-fixed HuT-78 (human Sezary syndrome cutaneous T lymphocyte cell line) cells labeling DGKZ/DGK-zeta with ab239081 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining in HuT-78 cell line.

Methanol is recommended for fixation as weak signal was observed using 4% PFA.

The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) (red) at 1/200 dilution.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody 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 (ab239081).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DGKZ/DGK-zeta antibody [EPR22040-80] - BSA and Azide free (AB239790)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling DGKZ/DGK-zeta with ab239081 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on subset of immune cells in human tonsil (PMID : 24573202) 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) Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab239081).

Perform heat mediated antigen retrieval 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-DGKZ/DGK-zeta antibody [EPR22040-80] - BSA and Azide free (AB239790)

Immunohistochemical analysis of paraffin-embedded rat cerebellum tissue labeling DGKZ/DGK-zeta with ab239081 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining on Purkinje cells of rat cerebellum (PMID : 14511325; PMID : 24119575) 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) Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab239081).

Perform heat mediated antigen retrieval 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-DGKZ/DGK-zeta antibody [EPR22040-80] - BSA and Azide free (AB239790)

Immunohistochemical analysis of paraffin-embedded mouse cerebellum tissue labeling DGKZ/DGK-zeta with ab239081 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic and nuclear staining on Purkinje cells of mouse cerebellum (PMID : 14511325; PMID : 24119575) 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) Ready to use.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab239081).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.