Anti-AK2 antibody [EPR11388(B)] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-AK2 antibody [EPR11388(B)] - BSA and Azide free (AB249383)

This data was developed using ab166901, the same antibody clone in a different buffer formulation.

Immunocytochemistry/Immunofluorescence analysis MCF-7 (human breast carcinoma) cells labelling AK2 with purified ab166901 at a dilution of 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised with 0.1% tritonX-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at dilution of 1/1000 was used as the secondary antibody. Nuclei counterstained with DAPI (blue).

Secondary Only Control : PBS was used instead of the primary antibody as the negative control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AK2 antibody [EPR11388(B)] - BSA and Azide free (AB249383)

This data was developed using ab166901, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human stomach tissue labeling AK2 using ab166901 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AK2 antibody [EPR11388(B)] - BSA and Azide free (AB249383)

This data was developed using ab166901, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling AK2 using ab166901 at 1/100 dilution.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IP

Unknown

Immunoprecipitation - Anti-AK2 antibody [EPR11388(B)] - BSA and Azide free (AB249383)

This data was developed using ab166901, the same antibody clone in a different buffer formulation.Immunoprecipitation of AK2 from HeLa cell lysate pellet using ab166901 at 1/10 dilution followed by immunoblotting. HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.

All lanes:

Immunoprecipitation - Anti-AK2 antibody [EPR11388(B)] (<a href='/en-us/products/primary-antibodies/ak2-antibody-epr11388b-ab166901'>ab166901</a>)

Predicted band size: 26 kDa

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• WB

Unknown

Western blot - Anti-AK2 antibody [EPR11388(B)] - BSA and Azide free (AB249383)

This data was developed using the same antibody clone in a different buffer formulation (ab166901).

Lanes 1 - 4 : Merged signal (red and green). Green - ab166901 observed at 26 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

ab166901 was shown to react with AK2 in wild-type HEK-293T cells in western blot with loss of signal observed in AK2 knockout cell line ab266539 (AK2 knockout cell lysate ab257825). Wild-type and AK2 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab166901 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-AK2 antibody [EPR11388(B)] (<a href='/en-us/products/primary-antibodies/ak2-antibody-epr11388b-ab166901'>ab166901</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

AK2 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human AK2 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-ak2-knockout-hek-293t-cell-line-ab266539'>ab266539</a>)

Lane 3:

HL-60 cell lysate at 20 µg

Lane 4:

HepG2 cell lysate at 20 µg

Predicted band size: 26 kDa

Observed band size: 26 kDa

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• WB

Unknown

Western blot - Anti-AK2 antibody [EPR11388(B)] - BSA and Azide free (AB249383)

This data was developed using ab166901, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-AK2 antibody [EPR11388(B)] (<a href='/en-us/products/primary-antibodies/ak2-antibody-epr11388b-ab166901'>ab166901</a>) at 1/1000 dilution

Lane 1:

Fetal kidney lysate at 10 µg

Lane 2:

Fetal liver lysate at 10 µg

Lane 3:

HepG2 cell lysate at 10 µg

Lane 4:

HeLa cell lysate at 10 µg

Secondary

All lanes:

goat anti-rabbit HRP antibody at 1/2000 dilution

Predicted band size: 26 kDa

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