Anti-ADK antibody [EPR23166-143]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADK antibody [EPR23166-143] (AB243636)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling ADK with ab243636 at 1/1000 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear and cytoplasmic staining on human liver is observed. The section was incubated with ab243636 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : 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 solution 2) for 20mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADK antibody [EPR23166-143] (AB243636)

Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling ADK with ab243636 at 1/1000 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear and weak cytoplasmic staining on human tonsil is observed. The section was incubated with ab243636 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : 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 solution 2) for 20mins.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-ADK antibody [EPR23166-143] (AB243636)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PC-3 (Human prostate adenocarcinoma epithelial cell) cells labeling ADK with ab243636 at 1\500 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 was used as the secondary antibody.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADK antibody [EPR23166-143] (AB243636)

Immunohistochemical analysis of paraffin-embedded mouse cerebrum tissue labeling ADK with ab243636 at 1/1000 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear and cytoplasmic staining on mouse cerebrum is observed. The section was incubated with ab243636 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : 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 solution 2) for 20mins.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ADK antibody [EPR23166-143] (AB243636)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling ADK with ab243636 at 1/1000 followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear and cytoplasmic staining on rat kidney is observed. The section was incubated with ab243636 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with hematoxylin.

Secondary antibody only control : 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 solution 2) for 20mins.

• WB

Lab

Western blot - Anti-ADK antibody [EPR23166-143] (AB243636)

Lanes 1- 2 : Merged signal (red and green). Green - ab243636 observed at 41 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab243636 was shown to react with Adenosine kinase in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265646 (knockout cell lysate ab257343) was used. Wild-type HeLa and ADK knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab243636 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 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 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-ADK antibody [EPR23166-143] (ab243636) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ADK knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ADK knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-adk-knockout-hela-cell-line-ab265646'>ab265646</a>)

Predicted band size: 41 kDa

Observed band size: 41 kDa

false

• WB

Lab

Western blot - Anti-ADK antibody [EPR23166-143] (AB243636)

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 29038540, 30944910).

All lanes:

Western blot - Anti-ADK antibody [EPR23166-143] (ab243636) at 1/1000 dilution

Lane 1:

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

Lane 2:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

Wild-type HAP1 whole cell lysate at 40 µg

Lane 4:

ADK knockout HAP1 whole cell lysate at 40 µg

Lane 5:

Hepa1-6 (mouse hepatoma epithelial cell), whole cell lysate at 20 µg

Lane 6:

PC-3 (human prostate adenocarcinoma epithelial cell), whole cell 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: 41 kDa

false

• WB

Lab

Western blot - Anti-ADK antibody [EPR23166-143] (AB243636)

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

The lower band (38 kDa) in liver lysates represents ADK-short isoform.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 29038540, 30944910).

Exposure time : Lanes 1-4 : 10 seconds; Lanes 5-6 : 26 seconds.

All lanes:

Western blot - Anti-ADK antibody [EPR23166-143] (ab243636) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell), whole cell lysate at 20 µg

Lane 2:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 20 µg

Lane 5:

Mouse liver lysate at 20 µg

Lane 6:

Rat liver 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: 41 kDa

Observed band size: 41 kDa

false