Anti-Gephyrin antibody [EPR12650] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Gephyrin antibody [EPR12650] - BSA and Azide free (AB250503)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling Gephyrin with purified ab181382 at 1/100 dilution (1.62 µg/ml). Heat mediated antigen retrieval was performed Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab181382)

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Gephyrin antibody [EPR12650] - BSA and Azide free (AB250503)

This data was developed using ab181382, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of parafiin embedded human brain tissue using ab181382 (unpurified).

Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

Immunohistochemical analysis of parafiin embedded human brain tissue using ab181382 (unpurified).

Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-Gephyrin antibody [EPR12650] - BSA and Azide free (AB250503)

This data was developed using ab181382, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Gephyrin antibody [EPR12650] - BSA and Azide free (ab250503) at 1/1000 dilution

Lane 1:

HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates at 20 µg

Lane 2:

Mouse heart lysates at 20 µg

Lane 3:

Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysates at 20 µg

Lane 4:

C6 (Rat glial tumor glial cell) whole cell lysates 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: 80 kDa

Observed band size: 93 kDa

false

• WB

Unknown

Western blot - Anti-Gephyrin antibody [EPR12650] - BSA and Azide free (AB250503)

This data was developed using ab181382, the same antibody clone in a different buffer formulation.

Lanes 1 - 4 : Merged signal (red and green). Green - ab181382 observed at 90 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab181382 was shown to recognize Gephyrin in wild-type HAP1 cells as signal was lost at the expected MW in GPHN (Gephyrin) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GPHN (Gephyrin) knockout samples were subjected to SDS-PAGE. ab181382 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 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 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Gephyrin antibody [EPR12650] - BSA and Azide free (ab250503) at 1 µg/mL

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

GPHN (Gephyrin) knockout HAP1 whole cell lysate at 20 µg

Lane 3:

MCF7 whole cell lysate at 20 µg

Lane 4:

U2OS whole cell lysate at 20 µg

Predicted band size: 80 kDa

false

• WB

Supplier Data

Western blot - Anti-Gephyrin antibody [EPR12650] - BSA and Azide free (AB250503)

This data was developed using ab181382, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-Gephyrin antibody [EPR12650] - BSA and Azide free (ab250503) at 1/1000 dilution

Lane 1:

Fetal brain tissue lysate at 10 µg

Lane 2:

293T cell lysate at 10 µg

Lane 3:

SH-SY5Y cell lysate at 10 µg

Predicted band size: 80 kDa

false