Anti-ZDHHC7 antibody [EPR30883-532]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ZDHHC7 antibody [EPR30883-532] (AB327351)

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling ZDHHC7 with ab327351 at 1/500 (0.954 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil. The primary antibody was incubated for 30 mins at room temperature. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ZDHHC7 antibody [EPR30883-532] (AB327351)

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling ZDHHC7 with ab327351 at 1/500 (0.954 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon cancer. The primary antibody was incubated for 30 mins at room temperature. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• mIHC

Lab

Multiplex immunohistochemistry - Anti-ZDHHC7 antibody [EPR30883-532] (AB327351)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human colon cancer tissue staining ZDHHC7 with ab327351 at a 1 : 200 (2.385 μg/ml) dilution, ab52649 anti-GM130 used at 1 : 100 (1.18 μg/ml) dilution.

Panel A : anti-ZDHHC7 (green; Opal™520), anti-GM130 (magenta; Opal™690) on human colon cancer.
Panel B : anti-ZDHHC7 showed Golgi staining in human colon cancer.
Panel C : anti-GM130 showed Golgi staining in human colon cancer.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab327351 and ab52649 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

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.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-ZDHHC7 antibody [EPR30883-532] (AB327351)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling ZDHHC7 with ab327351 at 1/50 (9.54 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing golgi staining in HepG2 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab169276 Anti-GM130 mouse polyclonal antibody - cis-Golgi Marker was used to counterstain GM130 at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-ZDHHC7 antibody [EPR30883-532] (AB327351)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Parental HeLa (human cervical adenocarcinoma epithelial cell) and ZDHHC7 knockout HeLa cells labelling ZDHHC7 with ab327351 at 1/50 (9.54 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing golgi staining in parental HeLa cells and no staining in ZDHHC7 knockout HeLa cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab169276 Anti-GM130 mouse polyclonal antibody - cis-Golgi Marker was used to counterstain GM130 at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• mIHC

Lab

Multiplex immunohistochemistry - Anti-ZDHHC7 antibody [EPR30883-532] (AB327351)

Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded human colon tissue staining ZDHHC7 with ab327351 at a 1 : 200 (2.385 μg/ml) dilution, ab52649 anti-GM130 used at 1 : 100 (1.18 μg/ml) dilution.

Panel A : anti-ZDHHC7 (green; Opal™520), anti-GM130 (magenta; Opal™690) on human colon.
Panel B : anti-ZDHHC7 showed Golgi staining in human colon.
Panel C : anti-GM130 showed Golgi staining in human colon.
Panel D : Nuclear DNA was labeled with DAPI (shown in blue).

The section was incubated in three rounds of staining : in the order of ab327351 and ab52649 for 30 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

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.

Nuclear counter stain with DAPI.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ZDHHC7 antibody [EPR30883-532] (AB327351)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling ZDHHC7 with ab327351 at 1/500 (0.954 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon. The primary antibody was incubated for 30 mins at room temperature. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• I-ELISA

Lab

Indirect ELISA - Anti-ZDHHC7 antibody [EPR30883-532] (AB327351)

Indirect ELISA analysis of ab327351 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution dilution.

Antigen : Human ZDHHC7; Human ZDHHC3.

Antigen concentration : 1000 ng/ml

This antibody does not cross-react with Human ZDHHC3 by ELISA.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-ZDHHC7 antibody [EPR30883-532] (AB327351)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) and NIH/3T3 (mouse embryonic fibroblast) cells labelling ZDHHC7 with ab327351 at 1/50 (9.54 μg/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing golgi staining in Neuro-2a cells and no staining in NIH/3T3 cells (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue).
Low expression : NIH/3T3.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab169276 Anti-GM130 mouse polyclonal antibody - cis-Golgi Marker was used to counterstain GM130 at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ZDHHC7 antibody [EPR30883-532] (AB327351)

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling ZDHHC7 with ab327351 at 1/500 (0.954 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen. The primary antibody was incubated for 30 mins at room temperature. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ZDHHC7 antibody [EPR30883-532] (AB327351)

Immunohistochemical analysis of paraffin-embedded Mouse T cell lymphoma tissue labeling ZDHHC7 with ab327351 at 1/500 (0.954 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse T cell lymphoma. The primary antibody was incubated for 30 mins at room temperature. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ZDHHC7 antibody [EPR30883-532] (AB327351)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling ZDHHC7 with ab327351 at 1/500 (0.954 μg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse colon. The primary antibody was incubated for 30 mins at room temperature. Incubate slides with 3% Hydrogen Peroxide for 10 mins at room temperature after secondary antibody incubation to reduce the background.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Lab

Western blot - Anti-ZDHHC7 antibody [EPR30883-532] (AB327351)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The identity of the higher MW band at approximately 75 kDa is unknown.

The samples are unboiled.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-ZDHHC7 antibody [EPR30883-532] (ab327351) at 1/1000 dilution

Lane 1:

Human colon tissue lysate at 40 µg

Lane 2:

Human testis tissue lysate at 40 µg

Lane 3:

Human breast cancer tissue lysate at 40 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 28 kDa,36 kDa

false

Exposure time: 26s

• WB

Lab

Western blot - Anti-ZDHHC7 antibody [EPR30883-532] (AB327351)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : NIH/3T3, spleen

The identity of the higher MW band at approximately 75 kDa is unknown.

The samples are unboiled.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-ZDHHC7 antibody [EPR30883-532] (ab327351) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 40 µg

Lane 2:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 40 µg

Lane 3:

786-O (human kidney epithelial cell) whole cell lysate at 40 µg

Lane 4:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 5:

Mouse colon tissue lysate at 40 µg

Lane 6:

Mouse spleen tissue lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 28 kDa,36 kDa

false

Exposure time: 26s

• WB

Lab

Western blot - Anti-ZDHHC7 antibody [EPR30883-532] (AB327351)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, ab327351 was shown to bind specifically to ZDHHC7. Target of interest was observed at 28 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in ZDHHC7 HeLa knockout cell lysates(lane 2).

The samples are unboiled.

The identity of the higher MW band at approximately 75 kDa is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

Exposure time : Lanes1-2 : 59 seconds, lanes 3-4 : 26 seconds

All lanes:

Western blot - Anti-ZDHHC7 antibody [EPR30883-532] (ab327351) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellular carcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 80 µg

Lane 2:

HepG2 transfected with siRNA specifically targeting ZDHHC7 whole cell lysate at 80 µg

Lane 3:

Wild-type HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 80 µg

Lane 4:

ZDHHC7 knockout HeLa whole cell lysate at 80 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 28 kDa,36 kDa

false