Anti-ALK antibody [EPR29054-38]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALK antibody [EPR29054-38] (AB323643)

Immunohistochemical analysis of paraffin-embedded (A) KARPAS-299 (human T cell lymphoma cell) cell pellet. (B) U-937 (human histiocytic lymphoma monocyte) cell pellet. tissue labeling ALK with ab323643 at 1/200 (2.63 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on (A) KARPAS-299 cell pellet, no staining on (B) U-937 cell pellet.
The section was incubated with ab323643 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 was performed 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-ALK antibody [EPR29054-38] (AB323643)

Immunohistochemical analysis of paraffin-embedded E15.5 Mouse embryo tissue labeling ALK with ab323643 at 1/100 (5.26 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on brain and peripheral nervous system of E15.5 mouse embryo.
The section was incubated with ab323643 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 was performed 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-ALK antibody [EPR29054-38] (AB323643)

Immunohistochemical analysis of paraffin-embedded (A) Mouse PDX model of human anaplastic large cell lymphoma. (B) Control mouse PDX model. tissue labeling ALK with ab323643 at 1/2000 (0.263 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on (A) Mouse PDX model of human anaplastic large cell lymphoma. No staining on (B) Control mouse PDX model.
The section was incubated with ab323643 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 was performed 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-ALK antibody [EPR29054-38] (AB323643)

Immunohistochemical analysis of paraffin-embedded (A) Neuro-2a (mouse neuroblastoma neuroblast) cell pellet. (B) RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cell pellet. tissue labeling ALK with ab323643 at 1/100 (5.26 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on (A) Neuro-2a cell pellet, no staining on (B) RAW 264.7 cell pellet.
The section was incubated with ab323643 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-ALK antibody [EPR29054-38] (AB323643)

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

Negative control : Raji (PMID : 30541790), Jurkat (PMID : 10793082).

Full-length ALK is approximately 250 kDa. Band around 130kDa might be cleavage fragment(PMID : 30541790).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 9736036, PMID : 11310834, 9121481).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-ALK antibody [EPR29054-38] (ab323643) at 1/1000 dilution

Lane 1:

IMR-32 (human neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 2:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) 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

Observed band size: 250 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-ALK antibody [EPR29054-38] (AB323643)

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

Negative control : U-937 (PMID : 9736036).

KARPAS-299 cells carry the NPM-ALK fusion (PMID : 9736036, PMID : 11310834). Bands higher than 250 KDa are likely representing oligomers of NPM-ALK (PMID : 23610619).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 9736036, PMID : 11310834, 9121481).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-ALK antibody [EPR29054-38] (ab323643) at 1/1000 dilution

Lane 1:

KARPAS-299 (human T cell lymphoma cell) whole cell lysate at 20 µg

Lane 2:

U-937 (human histiocytic lymphoma monocyte) 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

Observed band size: 80 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-ALK antibody [EPR29054-38] (AB323643)

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

Negative control : RAW 264.7 (30541790), kidney (PMID : 9053841).

Low expression : hippocampus (PMID : 37291945, PMID : 9053841).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 9736036, PMID : 11310834, 9121481).

The identity of the lower MW band at approximately 37 kDa (in lanes 4-5) is unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-ALK antibody [EPR29054-38] (ab323643) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 2:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 3:

Mouse P5 brain tissue lysate at 20 µg

Lane 4:

Mouse kidney tissue lysate at 20 µg

Lane 5:

Mouse hippocampus 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

Observed band size: 250 kDa,36 kDa

false

Exposure time: 180s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ALK antibody [EPR29054-38] (AB323643)

Immunohistochemical analysis of paraffin-embedded Adult mouse kidney tissue labeling ALK with ab323643 at 1/100 (5.26 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control : No staining on Adult mouse kidney.
The section was incubated with ab323643 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 was performed 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-ALK antibody [EPR29054-38] (AB323643)

Immunohistochemical analysis of paraffin-embedded Adult mouse cerebral cortex tissue labeling ALK with ab323643 at 1/100 (5.26 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : Nearly no staining on adult mouse cerebral cortex.
The section was incubated with ab323643 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 was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins