Anti-DNAJC6 antibody [EPR25332-124]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-DNAJC6 antibody [EPR25332-124] (AB289711)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized K-562 (human chronic myelogenous leukemia lymphoblast cell line) cells labeling DNAJC6 with ab289711 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081), at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC6 antibody [EPR25332-124] (AB289711)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling DNAJC6 with ab289711 at 1/4000(0.114ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : no staining on human liver (PMID : 25446072). The section was incubated with ab289711 for 30 mins at room temperature. 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC6 antibody [EPR25332-124] (AB289711)

Immunohistochemical analysis of paraffin-embedded Human brain tissue labelling DNAJC6 with ab289711 at 1/4000(0.114ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human cerebrum. The section was incubated with ab289711 for 30 mins at room temperature. 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC6 antibody [EPR25332-124] (AB289711)

Immunohistochemical analysis of paraffin-embedded Mouse brain tissue labelling DNAJC6 with ab289711 at 1/4000(0.114ug/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse cerebrum. The section was incubated with ab289711 for 30 mins at room temperature. 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-DNAJC6 antibody [EPR25332-124] (AB289711)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling DNAJC6 with ab289711 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081), at 1/2000 dilution was used as the secondary antibody.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (AB289711)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse liver (fresh) tissue labelling DNAJC6 with ab289711 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution (Green). Negative control (PMID : 23211418). No staining on mouse liver. The nuclear conterstain is DAPI (Blue).

Secondary antibody only control : Used PBS instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (AB289711)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse cortex (fresh) tissue labelling DNAJC6 with ab289711 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution (Green). Positive staining on mouse cortex. The nuclear conterstain is DAPI (Blue).

Secondary antibody only control : Used PBS instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (AB289711)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat liver (fresh) tissue labelling DNAJC6 with ab289711 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution (Green). Negative control (PMID : 23211418). No staining on rat liver. The nuclear conterstain is DAPI (Blue).

Secondary antibody only control : Used PBS instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-DNAJC6 antibody [EPR25332-124] (AB289711)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen rat cortex (fresh) tissue labelling DNAJC6 with ab289711 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution (Green). Positive staining on rat cortex. The nuclear conterstain is DAPI (Blue).

Secondary antibody only control : Used PBS instead of primary antibody, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) secondary antibody at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DNAJC6 antibody [EPR25332-124] (AB289711)

Immunohistochemical analysis of paraffin-embedded Rat brain tissue labelling DNAJC6 with ab289711 at 1/4000(0.114 µg/ml) dilution followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat cerebrum. The section was incubated with ab289711 for 30 mins at room temperature. 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 with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• WB

Lab

Western blot - Anti-DNAJC6 antibody [EPR25332-124] (AB289711)

Negative control : Liver, Skeletal muscle (PMID : 23211418)
The molecular weight observed is consistent with what has been described in the literature (PMID : 1973169)

Exposure times :

Lane 1-7 : 7.75 seconds
Lane 8-9 : 37 seconds

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

All lanes:

Western blot - Anti-DNAJC6 antibody [EPR25332-124] (ab289711) at 1/1000 dilution

Lane 1:

Human spinal cord tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Lane 4:

Mouse cerebral cortex tissue lysate at 20 µg

Lane 5:

Mouse brain tissue lysate at 20 µg

Lane 6:

Mouse liver tissue lysate at 20 µg

Lane 7:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 8:

Rat brain tissue lysate at 20 µg

Lane 9:

Rat liver tissue lysate at 20 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 1/100000 dilution

Predicted band size: 100 kDa

Observed band size: 110 kDa

false

• WB

Lab

Western blot - Anti-DNAJC6 antibody [EPR25332-124] (AB289711)

Fresh lysates were used in this WB.
The molecular weight observed is consistent with what has been described in the literature (PMID : 1973169).

5% NFDM/TBST was used as a blocking and diluting buffer.

All lanes:

Western blot - Anti-DNAJC6 antibody [EPR25332-124] (ab289711) at 1/1000 dilution

Lane 1:

U-87 MG (human glioblastoma-astrocytoma epithelial cell), whole cell lysate at 10 µg

Lane 2:

K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 10 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 1/100000 dilution

Predicted band size: 100 kDa

Observed band size: 110 kDa

false

Exposure time: 48s