Anti-IRF2 antibody [EPR4644(2)]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-IRF2 antibody [EPR4644(2)] (AB124744)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling IRF2 with ab124744 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Confocal image showing nuclear staining on HeLa cells.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows :
-ve control 1 : ab124744 at 1/500 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (AB124744)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cervical cancer tissue labeling IRF2 with purified ab124744 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution was used as the secondary antibody. Nucleus staining on tumor cells of human cervix cancer was observed. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (AB124744)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human colon tissue labeling IRF2 with purified ab124744 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution was used as the secondary antibody. Nucleus staining on epithelium of human colon was observed. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (AB124744)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colonic carcinoma tissue labelling IRF2 with purified ab124744 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRF2 antibody [EPR4644(2)] (AB124744)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse colon tissue labeling IRF2 with purified ab124744 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution was used as the secondary antibody. Nucleus staining on epithelial cells of mouse colon was observed. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.

• WB

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Western blot - Anti-IRF2 antibody [EPR4644(2)] (AB124744)

Blocking and Diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744) at 1/5000 dilution

Lane 1:

SW480 (Human colorectal adenocarcinoma cell line) at 20 µg

Lane 2:

HeLa (Human epithelial cells from cervix adenocarcinoma) at 20 µg

Lane 3:

Jurkat (Human T cell leukemia cells from peripheral blood) at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 39 kDa

Observed band size: 50 kDa

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Exposure time: 3min

• WB

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Western blot - Anti-IRF2 antibody [EPR4644(2)] (AB124744)

Lane 1 : Wild type HAP1 whole cell lysate (20 μg)
Lane 2 : IRF2 knockout HAP1 whole cell lysate (20 μg)
Lane 3 : HeLa whole cell lysate (20 μg)
Lane 4 : CACO2 whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab124744 observed at 48 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab124744 was shown to recognize IRF2 when IRF2 knockout samples were used, along with additional cross-reactive bands. Wild-type and IRF2 knockout samples were subjected to SDS-PAGE. ab124744 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 and 1/10000 dilutions respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) ab216773 and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744)

Predicted band size: 39 kDa

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• WB

Supplier Data

Western blot - Anti-IRF2 antibody [EPR4644(2)] (AB124744)

Blocking and Diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744) at 1/1000 dilution

Lane 1:

Caco-2 (Human colorectal adenocarcinoma cells) at 10 µg

Lane 2:

RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) at 10 µg

Lane 3:

NIH/3T3 (Mouse embyro fibroblast cells) at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 132 kDa,39 kDa

Observed band size: 132 kDa,50 kDa

false

Exposure time: 3min

• WB

Supplier Data

Western blot - Anti-IRF2 antibody [EPR4644(2)] (AB124744)

Blocking and Diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-IRF2 antibody [EPR4644(2)] (ab124744) at 1/1000 dilution

All lanes:

Human fetal lung lysate at 10 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 32 kDa,39 kDa

Observed band size: 32 kDa,50 kDa

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Exposure time: 3min

• OI-RD Scanning

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OI-RD Scanning - Anti-IRF2 antibody [EPR4644(2)] (AB124744)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.

• WB

CiteAb

Western blot - Anti-IRF2 antibody [EPR4644(2)] (AB124744)

IRF2 western blot using anti-IRF2 antibody [EPR4644(2)] ab124744. Publication image and figure legend from Cheng, L., Geng, L., et al., 2018, Cell Death Dis, PubMed 29695839.

ab124744 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 ab124744 please see the product overview.

There was an inverse correlation between expression of let-7a cluster and IRF-1 in the inflamed livers.a ELISA assay was used to detect the expression of such pro-inflammatory factors as IFN-γ, TNF-α, and IL-1β in liver specimens and corresponding serum sampled from the mice (n = 6) with hepatitis. We found that some slight differences in the expression of those three factors in liver specimens without reaching statistical significance. However, the expression of IFN-γ (p < 0.01), TNF-α (p < 0.05), and IL-1β (p < 0.01) were significantly increased in the serum of mice with hepatitis, particularly IFN-γ (fold change = 6.55). b Total RNA was prepared from HT29 human colon carcinoma cells and CT26.WT mouse colon carcinoma cells which had been treated with different concentrations of IFN-γ. The qTR-PCR results showed that treatment with IFN-γ increased IRF-1 expression at the mRNA level in both cell lines, whereas IRF-2 expression did not show a noticeable increase. c Total proteins were extracted from the pre-treated CRC cells and analyzed in western blot studies that used antibodies against IRF-1 and IRF-2. Histone was used as a loading control. c The differential expression of IRF-1 and IRF-2 in the liver specimens was also tested using IHC methods. The levels of IRF-1, but not IRF-2, were dramatically increased in the inflamed liver specimens, and IRF-1-positive structures were located mainly in regions where metastatic tumors were present. e Results from qRT-PCR studies showed that let-7a cluster expression was dramatically downregulated in both CRC cell lines after the cells had been treated with different concentrations of IFN-γ for 48 h (p < 0.05). A two-tailed t-test was used to compare data obtained from the qRT-PCR analyses of two independent groups. Values are shown as the mean ± SD. *p < 0.05; **p < 0.001

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