Anti-IRF3 (phospho S386) antibody [EPR2346]

• WB

Unknown

Western blot - Anti-IRF3 (phospho S386) antibody [EPR2346] (AB76493)

Blocking and diluting buffer : 5% NFDM/TBST

We are unsure to define the extra band at 37KD.

All lanes:

Western blot - Anti-IRF3 (phospho S386) antibody [EPR2346] (ab76493) at 0.2 µg/mL

Lane 1:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Lane 2:

MCF7 (Human breast adenocarcinoma epithelial cell) treated with calyculin A for 45 minutes whole cell lysates at 15 µg

Lane 3:

MCF7 (Human breast adenocarcinoma epithelial cell) treated with calyculin A for 45 minutes whole cell lysates. Then the membrane was incubated with phosphatase 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 1/20000 dilution

Predicted band size: 47 kDa

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• WB

Unknown

Western blot - Anti-IRF3 (phospho S386) antibody [EPR2346] (AB76493)

Blocking buffer - 5% NFDM/TBST

Diluting buffer - 1% BSA

All lanes:

Western blot - Anti-IRF3 (phospho S386) antibody [EPR2346] (ab76493) at 1/20000 dilution

Lane 1:

MCF7 cell lysate - treated with Calyculin A at 20 µg

Lane 2:

MCF7 cell lysate – untreated at 20 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 1/20000 dilution

Predicted band size: 47 kDa

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• Dot

Unknown

Dot Blot - Anti-IRF3 (phospho S386) antibody [EPR2346] (AB76493)

Dot blot analysis of IRF3 single phospho peptide pS386 (lane 1) and IRF3 non-phospho peptide (lane 2) with unpurified ab76493 at 1/1000. Blocking and diluting buffer was 5% NFDM/TBST. The secondary antibody used was ab97051 Peroxidase conjugated Goat Anti-Rabbit IgG, (H+L) at 1/100,000.

• WB

CiteAb

Western blot - Anti-IRF3 (phospho S386) antibody [EPR2346] (AB76493)

Western Blotting using Anti-IRF3 (phospho S386) antibody [EPR2346], ab76493. Publication image from Zhao, Y. et al., 2018, Nat Commun, 30451863. Legend direct from paper.

RLRs-MAVS signaling pathway restricts KSHV lytic reactivation in BC-3 cells. a RNA was extracted from BC-3 cells expressing the indicated shRNAs and the target mRNA level was quantified by RT-qPCR. b Expression of KSHV viral genes in cells described in (a) was quantified by RT- qPCR. c BC-3 F-RIG-I, F-MDA5, and Control (Con) cells were reactivated for 48 h. Cell lysates from latent and lytic cells were analyzed by immunoblotting for the indicated proteins. d RNA was extracted from BC-3 cells described in (c) and expression of the indicated viral genes was quantified by RT-qPCR. Error bars in all panels represent mean ± SD from three independent experiments. p-values were determined by the Student’s t test, *p < 0.05, **p < 0.01

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