Anti-Jarid2 antibody [EPR6357(2)] - BSA and Azide free

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Jarid2 antibody [EPR6357(2)] - BSA and Azide free (AB251123)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192252).

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% triton X-100 permeabilized SH-SY5Y cells labeling Jarid2 with ab192252 at 1/100 dilution, followed by Goat anti rabbit IgG (Alexa Fluor® 555) secondary antibody (ab150078) at 1/500 dilution. Nuclear counter stain Dapi (blue).

The two negative controls are ab192252 at 1/100 dilution followed by Goat anti mouse IgG (Alexa Fluor®488) secondary antibody at 1/200 dilution.

• ChIP

Unknown

ChIP - Anti-Jarid2 antibody [EPR6357(2)] - BSA and Azide free (AB251123)

Chromatin was prepared from HeLa cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
The ChIP was performed with 25 μg of chromatin, 5 μg of ab192252 (red), and 20 μl of Protein A/G sepharose beads. 5 μg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).

Primers and probes are located in the first kb of the transcribed region.
*http : //www.abcam.com/resources?keywords=X%20ChIP%20protocol

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192252).

• WB

Unknown

Western blot - Anti-Jarid2 antibody [EPR6357(2)] - BSA and Azide free (AB251123)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192252).

Blocking/Dilution buffer : 5% NFDM /TBST.

All lanes:

Western blot - Anti-Jarid2 antibody [EPR6357(2)] - BSA and Azide free (ab251123) at 1/10000 dilution

Lane 1:

NCCIT cell lysate at 20 µg

Lane 2:

SH-SY5Y cell lysate at 20 µg

Lane 3:

293 cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 138 kDa

Observed band size: 139 kDa

false

• WB

Unknown

Western blot - Anti-Jarid2 antibody [EPR6357(2)] - BSA and Azide free (AB251123)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192252).

Blocking/Dilution buffer : 5% NFDM /TBST.

All lanes:

Western blot - Anti-Jarid2 antibody [EPR6357(2)] - BSA and Azide free (ab251123) at 1/1000 dilution

All lanes:

U87-MG cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 138 kDa

false

• WB

Unknown

Western blot - Anti-Jarid2 antibody [EPR6357(2)] - BSA and Azide free (AB251123)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab192252).

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : JARID2 (KO) knockout HAP1 whole cell lysate (20 μg)
Lane 3 : SH-SY5Y whole cell lysate (20 μg)
Lane 4 : U87MG whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab192252 observed at 138 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab192252 was shown to specifically recognize JARID2 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when JARID2 knockout samples were examined. Wild-type and JARID2 knockout samples were subjected to SDS-PAGE. ab192252 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed 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 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Jarid2 antibody [EPR6357(2)] - BSA and Azide free (ab251123)

Predicted band size: 138 kDa

false