Anti-HPRT antibody [EPR5299]

• WB

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Western blot - Anti-HPRT antibody [EPR5299] (AB109021)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-HPRT antibody [EPR5299] (ab109021) at 0.02 µg/mL

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Lane 2:

Mouse brain lysates at 15 µg

Lane 3:

Rat brain lysates at 15 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 0.02 µg/mL

Predicted band size: 25 kDa

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• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HPRT antibody [EPR5299] (AB109021)

Unpurified ab109021 at 1/50 dilution staining HPRT in Human colon by Immunohistochemistry, Paraffin-embedded tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HPRT antibody [EPR5299] (AB109021)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human lung cancer tissue sections labeling HPRT with Purified ab109021 at 1 : 150 dilution (1.19 µg/ml). Heat mediated antigen retrieval was performed 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.Hematoxylinwas used as a counterstain

• IP

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Immunoprecipitation - Anti-HPRT antibody [EPR5299] (AB109021)

ab109021 (purified) at 1 : 30 dilution (2μg) immunoprecipitating HPRT in Rat brain lysate.
Lane 1 (input) : Rat brain lysate 10μg
Lane 2 (+) : ab109021 & Rat brain lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab109021 in Rat brain lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1 : 1000 dilution.
Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-HPRT antibody [EPR5299] (ab109021)

Predicted band size: 25 kDa

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• WB

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Western blot - Anti-HPRT antibody [EPR5299] (AB109021)

All lanes:

Western blot - Anti-HPRT antibody [EPR5299] (ab109021) at 1/10000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

HepG2 cell lysate at 10 µg

Lane 3:

A549 cell lysate at 10 µg

Lane 4:

MCF7 cell lysate at 10 µg

Predicted band size: 25 kDa

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• WB

Supplier Data

Western blot - Anti-HPRT antibody [EPR5299] (AB109021)

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 - ab109021 observed at 25 kDa. Red - loading control, ab8245, observed at 37 kDa.

Unpurified ab109021 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. ab109021 and ab8245 (Mouse anti GAPDH loading control) were both diluted at 1/10,000 and incubated overnight at 4°C. 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 [EPR5299] (ab109021)

Predicted band size: 25 kDa

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• WB

CiteAb

Western blot - Anti-HPRT antibody [EPR5299] (AB109021)

HPRT western blot using anti-HPRT antibody [EPR5299] ab109021. Publication image and figure legend from Mardani, I., Tomas Dalen, K., et al., 2019, Sci Rep, PubMed 31061399.

ab109021 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab109021 please see the product overview.

Regulation of Plin2. (A) Immunoblot analysis of Plin2 using protein lysates from control (refed) and O/N fasted C57Bl/6N mice hearts (n = 3). Full-length blots are presented in Supplementary Fig. S7. (B) Quantification of immunoblot analysis on Plin2 (n = 3). (C) Triglyceride content in hearts of control (refed) and overnight fasted mice (n = 3). (D) Confocal microscopy images on C57Bl/6N cardiomyocytes, shown as z-stack 1, 2, 3 and a max stack picture containing 30 z-stacks compromised into one image. Plin2 is shown in green, ORO (LDs) in red and nuclei in blue and co-localization of LDs and Plin2 in yellow (x630 magnification, scale bar : 10 μm). Data are presented as mean ± SEM, *p < 0.05 vs. Plin2+/+, **p < 0.01 vs. Plin2+/+. TG, triglycerides; O/N, overnight.

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