Anti-IDH1 抗体 [EPR12296]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IDH1 antibody [EPR12296] (AB172964)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human gastric cancer tissue sections labeling IDH1 with purified ab172964 at 1/200 dilution (0.60 µg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.

• IP

Lab

Immunoprecipitation - Anti-IDH1 antibody [EPR12296] (AB172964)

Purified ab172964 at 1/20 dilution (0.6μg) immunoprecipitating IDH1 in HepG2 whole cell lysate.
Lane 1 (input) : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate.
Lane 2 (+) : ab172964 + HepG2 whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab172964 in HepG2 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-IDH1 antibody [EPR12296] (ab172964)

Predicted band size: 46 kDa

Observed band size: 47 kDa

false

• WB

Lab

Western blot - Anti-IDH1 antibody [EPR12296] (AB172964)

This blot was produced using 4-20% SDS-PAGE containing 15 μg of Raji whole cell lysate per lane at 150V for 1hr before being transferred onto a 0.45 μm PVDF membrane at 75V for 1hr. The membrane was then blocked for 1hr using 5% NFDM/TBST, then incubated with ab172964 (1/2000) at room temperature for 1hr. After being washed three times in TBST, the membrane was incubated with Peroxidase conjugated goat anti-rabbit IgG (H+L) (ab97051) at 1/20,000 dilution for 1hr at room temperature. The membrane was washed three times again. Then the signal was developed using the ECL technique.

ab172964 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in WB. The image shows the band intensity remains relatively constant across all storage temperatures, demonstrating that antibody activity is not affected under these conditions. This data was generated as part of a structured antibody stability study supporting Abcam’s transition from cold‑chain to ambient shipping.

All lanes:

Western blot - Anti-IDH1 antibody [EPR12296] (ab172964) at 1/2000 dilution

All lanes:

Raji whole cell lysate at 15 µg with NDFM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-IDH1 antibody [EPR12296] (AB172964)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : IDH1 knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HeLa whole cell lysate (20 μg)
Lane 4 : HepG2 whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab172964 observed at 46 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab172964 was shown to specifically react with IDH1 in wild-type HAP1 cells. No band was observed when IDH1 knockout samples were used. Wild-type and IDH1 knockout samples were subjected to SDS-PAGE. ab172964 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-IDH1 antibody [EPR12296] (ab172964)

Predicted band size: 46 kDa

false

• WB

Unknown

Western blot - Anti-IDH1 antibody [EPR12296] (AB172964)

All lanes:

Western blot - Anti-IDH1 antibody [EPR12296] (ab172964) at 1/1000 dilution

All lanes:

Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 46 kDa

false

• WB

Unknown

Western blot - Anti-IDH1 antibody [EPR12296] (AB172964)

All lanes:

Western blot - Anti-IDH1 antibody [EPR12296] (ab172964) at 1/1000 dilution

Lane 1:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate

Lane 2:

Rat kidney lysate

Lane 3:

Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 46 kDa

false