Anti-Aldolase+Aldolase C antibody [EPR19355] - BSA and Azide free

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Aldolase+Aldolase C antibody [EPR19355] - BSA and Azide free (AB251296)

This data was developed using ab200049, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling Aldolase + Aldolase C with ab200049 at 1/600 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/500 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Aldolase+Aldolase C antibody [EPR19355] - BSA and Azide free (AB251296)

This data was developed using ab200049, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed U-87 MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling Aldolase C + Aldolase with ab200049 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on U-87 MG cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab200049 at 1/500 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2 : Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Aldolase+Aldolase C antibody [EPR19355] - BSA and Azide free (AB251296)

This data was developed using ab200049, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed NIH/3T3 (Mouse embryonic fibroblast cell line) cells labeling Aldolase C + Aldolase with ab200049 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab200049 at 1/500 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/1000 dilution.
-ve control 2 : Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-Aldolase+Aldolase C antibody [EPR19355] - BSA and Azide free (AB251296)

This data was developed using ab200049, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times : Lanes 1-3 : 8 seconds; Lanes 4-7 : 10 seconds.

The expression profile observed is consistent with what has been described in the literature (PMID : 24475166 and 17659271).

Lanes 1 - 3:

Western blot - Anti-Aldolase + Aldolase C antibody [EPR19355] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/aldolase-aldolase-c-antibody-epr19355-astrocyte-marker-ab200049'>ab200049</a>) at 1/5000 dilution

Lanes 4 - 7:

Western blot - Anti-Aldolase + Aldolase C antibody [EPR19355] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/aldolase-aldolase-c-antibody-epr19355-astrocyte-marker-ab200049'>ab200049</a>) at 1/2000 dilution

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg

Lane 2:

U 87 MG (Human glioblastoma-astrocytoma epithelial cell line) whole cell lysate at 10 µg

Lane 3:

Human cerebellum lysate at 10 µg

Lane 4:

Human fetal liver lysate at 10 µg

Lane 5:

Human fetal heart lysate at 10 µg

Lane 6:

Human fetal kidney lysate at 10 µg

Lane 7:

human fetal spleenlysate at 10 µg

Secondary

Lanes 1 - 3:

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

Lanes 4 - 7:

Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

Predicted band size: 39 kDa

Observed band size: 39 kDa

false

• WB

Supplier Data

Western blot - Anti-Aldolase+Aldolase C antibody [EPR19355] - BSA and Azide free (AB251296)

This data was developed using ab200049, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Human ALDOC full length recombinant protein contains aa2-364 with a His-Tag^®. Human ALDOA and ALDOB full length recombinant protein contains aa1-364 with a His-Tag^®. These three recombinant proteins were made in-house.

This product showed very weak reactivity to Aldolase B, relative to Aldolase and Aldolase C, as demonstrated by WB of recombinant proteins.

All lanes:

Western blot - Anti-Aldolase + Aldolase C antibody [EPR19355] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/aldolase-aldolase-c-antibody-epr19355-astrocyte-marker-ab200049'>ab200049</a>) at 1/1000 dilution

Lane 1:

Human Aldolase C full length recombinant protein at 0.01 µg

Lane 2:

Human Aldolase full length recombinant protein at 0.01 µg

Lane 3:

Human Aldolase B full length recombinant protein at 0.01 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: 39 kDa

Observed band size: 39 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-Aldolase+Aldolase C antibody [EPR19355] - BSA and Azide free (AB251296)

This data was developed using ab200049, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure times : Lanes 1-5 : 8 seconds; Lanes 6-7 : 3 seconds.

Lanes 1 - 6:

Western blot - Anti-Aldolase + Aldolase C antibody [EPR19355] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/aldolase-aldolase-c-antibody-epr19355-astrocyte-marker-ab200049'>ab200049</a>) at 1/5000 dilution

Lane 7:

Western blot - Anti-Aldolase + Aldolase C antibody [EPR19355] - Astrocyte Marker (<a href='/en-us/products/primary-antibodies/aldolase-aldolase-c-antibody-epr19355-astrocyte-marker-ab200049'>ab200049</a>) at 1/2000 dilution

Lane 1:

C6 (Rat glial tumor cell line) whole cell lysate at 10 µg

Lane 2:

Rat brain tissue lysate at 10 µg

Lane 3:

Rat kidney tissue lysate at 10 µg

Lane 4:

Mouse heart tissue lysate at 10 µg

Lane 5:

RAW 264.7 (Mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg

Lane 6:

Mouse brain tissue lysate at 10 µg

Lane 7:

NIH/3T3 (Mouse embryonic fibroblast cell line) 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: 39 kDa

Observed band size: 39 kDa

false