重组Anti-JMJD6抗体[EPR23672-71] - BSA and Azide free

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (AB282015)

This data was developed using ab256798, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized parental HEK293T (Human embryonic kidney epithelial cell, Right) cells / JMJD6 KO HEK293T cells (Left) labelling JMJD6 with ab256798 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody. Positive staining on 293T cells (ab255449), while no staining on JMJD6 knockout HEK-293T cells (ab266402).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (AB282015)

This data was developed using ab256798, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tumor tissue labelling JMJD6 with ab256798 at 1/500 dilution (1.152 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human liver tumor. The section was incubated with ab256798 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (AB282015)

This data was developed using ab256798, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human liver tissue labelling JMJD6 with ab256798 at 1/500 dilution (1.152 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Negative control : No staining on human liver. The section was incubated with ab256798 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (AB282015)

This data was developed using ab256798, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human testis tissue labelling JMJD6 with ab256798 at 1/500 dilution (1.152 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human testis. The section was incubated with ab256798 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• WB

Lab

Western blot - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (AB282015)

This data was developed using ab256798, the same antibody clone in a different buffer formulation.

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

Lysates were made freshly and used in WB test immediately to minimize protein degradation.

The molecular weight observed is consistent with what has been described in the literature (PMID : 24360279).

This antibody reacts with an unidentifiable protein around 38 kDa.

Exposure time : 48 seconds.

All lanes:

Western blot - Anti-JMJD6 antibody [EPR23672-71] (<a href='/products/primary-antibodies/jmjd6-antibody-epr23672-71-ab256798'>ab256798</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 46 kDa

Observed band size: 147 kDa,294 kDa,49 kDa

false

• WB

Lab

Western blot - Anti-JMJD6 antibody [EPR23672-71] - BSA and Azide free (AB282015)

This data was developed using ab256798, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : Intercept^® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

Lanes 1-5 : Merged signal (red and green).

Green - ab256798 observed at 49, 147, 294kDa.

Red - loading control ab8245 observed at 36 kDa.

ab256798 Anti-JMJD6 antibody [EPR23672-71] was shown to react with JMJD6 in Hela cells in Western blot. Loss of signal was observed when knockout cell line

ab266402 (JMJD6 knockout cell lysate ab257490) was used. Wild-type and JMJD6 knockout samples were subjected to SDS-PAGE.

ab256798 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

The molecular weight observed is consistent with what has been described in the literature (PMID : 24360279).

This antibody reacts with an unidentifiable protein around 38 kDa.

All lanes:

Western blot - Anti-JMJD6 antibody [EPR23672-71] (<a href='/products/primary-antibodies/jmjd6-antibody-epr23672-71-ab256798'>ab256798</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

JMJD6 knockout HEK-293T whole cell lysate at 20 µg

Lane 3:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 5:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/10000 dilution

Predicted band size: 46 kDa

Observed band size: 147 kDa,294 kDa,49 kDa

false