Anti-ATIC抗体[F38 P7 H9]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATIC antibody [F38 P7 H9] (AB33520)

ab33520, at a concentration of 5 μg/ml, staining ATIC in Human colon carcinoma tissue by Immunohistochemistry.

• Flow Cyt

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Flow Cytometry - Anti-ATIC antibody [F38 P7 H9] (AB33520)

Overlay histogram showing HeLa cells stained with ab33520 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab33520, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• WB

Supplier Data

Western blot - Anti-ATIC antibody [F38 P7 H9] (AB33520)

All lanes:

Western blot - Anti-ATIC antibody [F38 P7 H9] (ab33520) at 2 µg/mL

Lane 1:

Hep G2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 30 µg

Lane 2:

HCT116 (Human colorectal carcinoma cell line) whole cell lysate at 30 µg

Lane 3:

K-562 (Human chronic myelogenous leukemia lymphoblast cell line ) whole cell lysate at 30 µg

Lane 4:

COLO 205 (Human colon adenocarcinoma cell line) whole cell lysate at 30 µg

Lane 5:

NTERA-2 (Human malignant pluripotent embryonic carcinoma cell line) whole cell lysate at 30 µg

Lane 6:

HT-29 (Human colorectal adenocarcinoma cell line) whole cell lysate at 30 µg

Secondary

All lanes:

Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate at 1/2500 dilution

Predicted band size: 65 kDa

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• WB

CiteAb

Western blot - Anti-ATIC antibody [F38 P7 H9] (AB33520)

ATIC western blot using anti-ATIC antibody [F38 P7 H9] ab33520. Publication image and figure legend from Parviainen, V. I., Joenväärä, S., et al., 2013, Proteome Sci, PubMed 23915316.

ab33520 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 ab33520 please see the product overview.

Western blot validation. NME2, ATIC and RAB5 were analyzed with Western Blot to validate the MS results. The WB quantification displayed good correlation with MS results. Average NME2 induced/control ratio in MS was 0.55 and with WB 0.69. For ATIC, the ratios were 0.62 (MS) and 0.68 (WB) and for RAB5 the ratios were 0.66 (MS) and 0.69 (WB). Beta-actin was used as loading control. The Western Blot results were normalized to average control values. Bars represent the average of three biological repeats and the error bars represent lowest and highest WB quantification results of the three biological replicates.

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