Anti-IMPDH2 antibody [EPR8365(B)] - BSA and Azide free

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-IMPDH2 antibody [EPR8365(B)] - BSA and Azide free (AB248323)

This data was developed using ab129165, the same antibody clone in a different buffer formulation.

Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labelling IMPDH2 (Green) with ab129165 at 1/1000 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150081, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG H&L, was used as the secondary antibody at a 1/1000. Tubulin (Red) was stained with ab195889, an Alexa Fluor^® 594-conjugated anti-alpha tubulin antibody, at a 1/200 dilution. DAPI (Blue) was used as the nuclear counterstain. Negative control using only secondary antibody.

Confocal image showing cytoplasmic and weak nuclear staining in HeLa cells.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-IMPDH2 antibody [EPR8365(B)] - BSA and Azide free (AB248323)

This data was developed using ab129165, the same antibody clone in a different buffer formulation. Intracellular Flow Cytometry analysis of K-562 (human chronic myelogenous leukemia lymphoblast) cells labeling IMPDH2 with ab129165 at 1/20 dilution (Red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% Methanol. Rabbit monoclonal - Alexa Fluor^® 488 (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

• IHC-FoFr

Lab

Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-IMPDH2 antibody [EPR8365(B)] - BSA and Azide free (AB248323)

This data was developed using ab129165, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling IMPDH2 with ab129165 at 1/4000 for 30 mins at room temperature. A Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody.

Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. Negative control using only secondary antibody. Counterstained with hematoxylin.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-FoFr

Lab

Immunohistochemistry (PFA perfusion fixed frozen sections) - Anti-IMPDH2 antibody [EPR8365(B)] - BSA and Azide free (AB248323)

This data was developed using ab129165, the same antibody clone in a different buffer formulation.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovarian cancer tissue labelling IMPDH2 with ab129165 at 1/4000 for 30 mins at room temperature. A Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody.

Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. Negative control using only secondary antibody. Counterstained with hematoxylin.

The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• WB

Lab

Western blot - Anti-IMPDH2 antibody [EPR8365(B)] - BSA and Azide free (AB248323)

This data was developed using ab129165, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST

Lanes 1 - 6:

Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (<a href='/en-us/products/primary-antibodies/met-c-met-antibody-ep1454y-n-terminal-ab51067'>ab51067</a>) at 1/1000 dilution

Lanes 1 - 2:

Western blot - Anti-IMPDH2 antibody [EPR8365(B)] (<a href='/en-us/products/primary-antibodies/impdh2-antibody-epr8365b-ab129165'>ab129165</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

MET knockout A549 cell lysate at 20 µg

Lane 3:

Wild-type HeLa ab255929 cell lysate at 20 µg

Lane 4:

MET (Met (c-Met)) knockout HeLa <a href='/en-us/products/cell-lysates/human-met-c-met-knockout-hela-cell-lysate-ab256991'>ab256991</a> cell lysate at 20 µg

Lane 5:

HT-29 cell lysate at 20 µg

Lane 6:

T-47D cell lysate at 20 µg

Secondary

All lanes:

NFDM/TBST at 1/20000 dilution

Observed band size: 40-50 kDa

false

• WB

Lab

Western blot - Anti-IMPDH2 antibody [EPR8365(B)] - BSA and Azide free (AB248323)

This data was developed using ab129165, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-IMPDH2 antibody [EPR8365(B)] (<a href='/en-us/products/primary-antibodies/impdh2-antibody-epr8365b-ab129165'>ab129165</a>) at 1/10000 dilution

Lane 1:

K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 2:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lanes 3 - 4:

Rat spleen lysate at 20 µg

Lane 5:

Mouse spleen 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 1/20000 dilution

Predicted band size: 56 kDa

Observed band size: 56 kDa

false

Exposure time: 60s

• WB

Lab

Western blot - Anti-IMPDH2 antibody [EPR8365(B)] - BSA and Azide free (AB248323)

This data was developed using ab129165, the same antibody clone in a different buffer formulation.

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

All lanes:

Western blot - Anti-IMPDH2 antibody [EPR8365(B)] (<a href='/en-us/products/primary-antibodies/impdh2-antibody-epr8365b-ab129165'>ab129165</a>) at 1/1000 dilution

Lane 1:

DDDK tagged human full length IMPDH1 recombinant protein at 10 µg

Lane 2:

DDDK tagged human full length IMPDH2 recombinant protein at 10 µg

Secondary

All lanes:

NFDM/TBST at 1/20000 dilution

Predicted band size: 56 kDa

Observed band size: 56 kDa

false

Exposure time: 140s