Anti-IDH1 antibody [RcMab-1]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [RcMab-1] (AB256557)

Immunohistochemical analysis of paraffin-embedded human prostatic hyperplasia tissue labeling IDH1 with ab256557 at 1/500 dilution (14.79μg/ml) followed by ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882). Positive staining on human prostatic hyperplasia. The section was incubated with ab256557 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-IDH1 antibody [RcMab-1] (AB256557)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized wild-type HAP1 cells labelling IDH1 with ab256557 at 1/20 dilution (73.95μg/ml), followed by ab150157 Goat Anti-rat IgG H&L (Alexa Fluor^® 488) antibody at 1/1000 dilution (Green).

Confocal image showing positive staining in wild-type HAP1 cells, while no staining in IDH1 knockout HAP1 cells.

ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 dilution, followed by ab150080 Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 594) at a 1/1000 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only controls.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [RcMab-1] (AB256557)

Immunohistochemical analysis of paraffin-embedded human normal cerebrum tissue labeling IDH1 with ab256557 at 1/500 dilution (14.79μg/ml) followed by ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882). The section was incubated with ab256557 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Negative control : No staining on human normal cerebrum.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [RcMab-1] (AB256557)

Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling IDH1 with ab256557 at 1/500 dilution (14.79μg/ml) followed by ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882). Positive staining on human kidney tissue. The section was incubated with ab256557 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• WB

Lab

Western blot - Anti-IDH1 antibody [RcMab-1] (AB256557)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-IDH1 antibody [RcMab-1] (ab256557) at 1.479 µg/mL

Lane 1:

Wild type HAP1 whole cell lysate at 20 µg

Lane 2:

IDH1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 5:

Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/10000 dilution

Predicted band size: 46 kDa

false

Exposure time: 8s

• WB

Lab

Western blot - Anti-IDH1 antibody [RcMab-1] (AB256557)

Blocking/Dilution buffer : 5% NFDM/TBST.

Exposure times : Lane 1 : 3 minutes; Lanes 2-3 : 3 seconds.

All lanes:

Western blot - Anti-IDH1 antibody [RcMab-1] (ab256557) at 1.479 µg/mL

Lane 1:

PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg

Lane 2:

Rat heart tissue lysate at 10 µg

Lane 3:

Human heart tissue lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/10000 dilution

Predicted band size: 46 kDa

false