Anti-HPRT antibody

• WB

Lab

Western blot - Anti-HPRT antibody (AB10479)

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)
Lane 2 : HPRT1 knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HeLa whole cell lysate (20 μg)
Lane 4 : A431 whole cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab10479 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab10479 was shown to specifically react with HPRT1 in wild-type HAP1 cells. No band was observed when HPRT1 knockout samples were examined. Wild-type and HPRT1 knockout samples were subjected to SDS-PAGE. ab10479 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/500 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-HPRT antibody (ab10479)

Predicted band size: 25 kDa

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• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-HPRT antibody (AB10479)

ab10479 staining HPRT in HepG2 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab10479 at 5µg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• WB

Lab

Western blot - Anti-HPRT antibody (AB10479)

Lane 1 : HeLa whole cell lysate (20 µg) Lane 2 : NIH/3T3 whole cell lysate (20 µg) Lane 3 : PC12 whole cell lysate (20 µg) Lane 4 : Wild-type HAP1 whole cell lysate (20 µg) Lane 5 : Empty Lane Lane 6 : HPRT1 knockout HAP1 whole cell lysate (20 µg) Lanes 1 - 6 : Merged signal (red and green). Green - ab10479 observed at 24 kDa. Red - loading control ab6301 observed at 95 kDa. Gel type : MES Blocking buffer : LiCOR blocking buffer ab10479 was shown to specifically react with HPRT1 in wild-type HAP1 cells. No band was observed when HPRT1 knockout samples were examined. Wild-type and HPRT1 knockout samples were subjected to SDS-PAGE. ab10479 and ab6301 (Beta catenin - Loading control) were incubated overnight at 4°C at 1 µg/ml and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800RD) preadsorbed and Goat anti-Mouse IgG H&L (IRDye® 680CW) preadsorbed secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-HPRT antibody (ab10479) at 1 µg/mL

Lane 1:

HeLa whole cell lysate at 20 µg

Lane 2:

NIH/3T3 whole cell lysate at 20 µg

Lane 3:

PC12 whole cell lysate at 20 µg

Lane 4:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 6:

HPRT1 knockout HAP1 whole cell lysate at 20 µg

Secondary

Lanes 1 - 6:

Goat anti-Mouse IgG H&L (IRDye® 680CW) preadsorbed at 1/10000 dilution

Lanes 1 - 6:

Goat anti-Rabbit IgG H&L (IRDye® 800RD) preadsorbed at 1/10000 dilution

Predicted band size: 25 kDa

Observed band size: 24 kDa

false

• WB

Unknown

Western blot - Anti-HPRT antibody (AB10479)

All lanes:

Western blot - Anti-HPRT antibody (ab10479) at 1/500 dilution

All lanes:

Western blot - Recombinant Human HPRT protein (His tag N-Terminus) (<a href='/en-us/products/proteins-peptides/recombinant-human-hprt-protein-ab97411'>ab97411</a>) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 25 kDa

true

Exposure time: 1min

• WB

Unknown

Western blot - Anti-HPRT antibody (AB10479)

All lanes:

Western blot - Anti-HPRT antibody (ab10479) at 1/500 dilution

All lanes:

Western blot - Recombinant Human HPRT protein (His tag N-Terminus) (<a href='/en-us/products/proteins-peptides/recombinant-human-hprt-protein-ab97411'>ab97411</a>) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 25 kDa

true

Exposure time: 1min

• WB

Project

Western blot - Anti-HPRT antibody (AB10479)

All lanes:

Western blot - Anti-HPRT antibody (ab10479) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 2:

Western blot - A-431 whole cell lysate (<a href='/en-us/products/cell-lysates/a-431-whole-cell-lysate-ab7909'>ab7909</a>) at 20 µg

Lane 3:

MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 20 µg

Lane 4:

Western blot - HEK-293 whole cell lysate (<a href='/en-us/products/cell-lysates/hek-293-whole-cell-lysate-ab7902'>ab7902</a>) at 20 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG H&L (HRP) Pre-Adsorbed at 1/10000 dilution

Predicted band size: 25 kDa

Observed band size: 24 kDa

false

• WB

Project

Western blot - Anti-HPRT antibody (AB10479)

All lanes:

Western blot - Anti-HPRT antibody (ab10479) at 1 µg/mL

Lane 1:

Western blot - NIH/3T3 whole cell lysate (<a href='/en-us/products/cell-lysates/nih-3t3-whole-cell-lysate-ab7179'>ab7179</a>) at 10 µg

Lane 2:

MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 10 µg

Lane 3:

Brain (Mouse) Tissue Lysate at 10 µg

Secondary

All lanes:

IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

Predicted band size: 25 kDa

Observed band size: 24 kDa,28 kDa

false