重组Anti-RENT1/hUPF1抗体[EPR4681]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RENT1/hUPF1 antibody [EPR4681] (AB109363)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human liver tissue sections labeling RENT1/hUPF1 with Purified ab109363 at 1 : 100 dilution (5.14 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. Hematoxylin was used as a counterstain.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RENT1/hUPF1 antibody [EPR4681] (AB109363)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RENT1/hUPF1 with Purified ab109363 at 1 : 100 dilution (5.1 μg/ml). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1 : 200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 μg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-RENT1/hUPF1 antibody [EPR4681] (AB109363)

Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling RENT1/hUPF1 with Purified ab109363 at 1/50 dilution (10μg/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-RENT1/hUPF1 antibody [EPR4681] (AB109363)

Overlay histogram showing HeLa cells stained with ab109363 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab109363, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/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 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

This image was generated using the unpurified version of the product.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RENT1/hUPF1 antibody [EPR4681] (AB109363)

ab109363, at 1/100, staining RENT1/hUPF1 in Human transitional cell carcinoma tissue by immunohistochemistry.

This image was generated using the unpurified version of the product.

Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RENT1/hUPF1 antibody [EPR4681] (AB109363)

ab109363, at 1/100, staining RENT1/hUPF1 in Human kidney tissue by immunohistochemistry.

This image was generated using the unpurified version of the product.

Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-RENT1/hUPF1 antibody [EPR4681] (AB109363)

ab109363, at 1/100, staining RENT1/hUPF1 in HeLa cells by immunofluorescence.

This image was generated using the unpurified version of the product.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-RENT1/hUPF1 antibody [EPR4681] (AB109363)

Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling RENT1/hUPF1 with purified ab109363 at 1/500. Cells were fixed with 100% methanol. ab150077 Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Nuclei were counterstained with DAPI (blue).

Secondary Only Control : PBS was used instead of the primary antibody as the negative control

This image was generated using the unpurified version of the product.

• IP

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Immunoprecipitation - Anti-RENT1/hUPF1 antibody [EPR4681] (AB109363)

ab109363 (purified) at 1 : 30 dilution (2μg) immunoprecipitating RENT1/hUPF1 in Raji whole cell lysate.
Lane 1 (input) : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate 10μg
Lane 2 (+) : ab109363 & Raji whole cell lysate
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab109363 in Raji whole cell 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-RENT1/hUPF1 antibody [EPR4681] (ab109363)

Predicted band size: 124 kDa

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• WB

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Western blot - Anti-RENT1/hUPF1 antibody [EPR4681] (AB109363)

This image was generated using the unpurified version of the product.

All lanes:

Western blot - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363) at 1/10000 dilution

Lane 1:

HuT-78 cell lysate at 10 µg

Lane 2:

Raji cell lysate at 10 µg

Lane 3:

SH-SY5Y cell lysate at 10 µg

Lane 4:

HeLa cell lysate at 10 µg

Predicted band size: 124 kDa

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• WB

Lab

Western blot - Anti-RENT1/hUPF1 antibody [EPR4681] (AB109363)

All lanes:

Western blot - Anti-RENT1/hUPF1 antibody [EPR4681] (ab109363) at 1/10000 dilution

Lane 1:

HuT-78 (Human Sezary syndrome cutaneous T lymphocyte) whole cell lysates at 20 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates at 20 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: 124 kDa

Observed band size: 130 kDa

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• WB

CiteAb

Western blot - Anti-RENT1/hUPF1 antibody [EPR4681] (AB109363)

RENT1/hUPF1 western blot using anti-RENT1/hUPF1 antibody [EPR4681] ab109363. Publication image and figure legend from Mooney, C. M., Jiménez-Mateos, E. M., et al., 2017, Sci Rep, PubMed 28128343.

ab109363 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 ab109363 please see the product overview.

NMD proteins are increased after SE in mice and Upf1 is increased in human TLE samples.(a–d) Protein levels of Upf1, phosho-Upf1 (p-Upf1), Upf2 and Upf3b in the ipsilateral hippocampus in control (C) mice and at 1, 4, 8 and 24 hours (h) after SE were analysed by western blot and semi-quantified. (a) Upf1 levels significantly increased 8 and 24 h after SE (n = 4/group; ANOVA, Dunnett's posthoc test ***p < 0.001, **p < 0.01). (b) p-Upf1 levels significantly increased 8 and 24 h after SE (n = 5/group; ANOVA, Dunnett's posthoc test *p < 0.05 compared to control. (c) Upf2 levels were increased 8 h after SE (n = 4/group; ANOVA, Dunnett's posthoc test *p < 0.05 compared to control). (d) Upf3b levels did not change after SE (n = 4/group; ANOVA, Dunnett's posthoc test p = 0.88). (e) Upf1 protein levels were significantly higher in TLE patients with hippocampal sclerosis (HS) compared to post-mortem controls and TLE patients without HS (Controls n = 6; TLE without HS n = 3, TLE with HS n = 3; ANOVA with Bonferroni post-hoc test comparing all columns; p = 0.0072). Representative blots have been cropped to reduce unnecessary area.

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