HA750283

IRF3 Recombinant Rabbit Monoclonal Antibody [SD2062]

cat.: HA750283

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, FC, IP
Clonality:	Monoclonal
Clone number:	SD2062

Form:	Liquid
Storage condition:	Store at 2-8℃. Avoid freeze.
Storage buffer:	1*PBS (pH7.4).
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 47 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human IRF3 aa 71-120 / 427.
Positive control:	HeLa cell lysate, Jurkat cell lysate, MCF7 cell lysate, A549 cell lysate, THP-1 cell lysate, Hela-si NT cell lysate, Hela-si IRF3 cell lysate, RAW264.7 cell lysate, mouse spleen tissue lysate, mouse liver tissue lysate, Hela, MCF-7, PANC-1, human breast carcinoma tissue, human kidney tissue, human pancreas tissue, human tonsil tissue, mouse colon tissue, mouse testis tissue, mouse heart tissue, rat brain tissue, rat colon tissue, rat heart tissue, rat spleen tissue.
Subcellular location:	Cytoplasm, Nucleus.
Uniprot #:	SwissProt: Q14653 Human \| P70671 Mouse
Alternative names:	IIAE7 Interferon regulatory factor 3 IRF 3 IRF-3 IRF3 IRF3_HUMAN MGC94729

Images

Fig1: This data was developed using HA750283, the same antibody clone in a different buffer formulation.
Western blot analysis of IRF3 on different lysates with Rabbit anti-IRF3 antibody (HA750283) at 1/5,000 dilution.

Lane 1: HeLa cell lysate (15 µg/Lane)
Lane 2: Jurkat cell lysate (15 µg/Lane)
Lane 3: MCF7 cell lysate (15 µg/Lane)
Lane 4: A549 cell lysate (15 µg/Lane)
Lane 5: THP-1 cell lysate (15 µg/Lane)

Predicted band size: 47 kDa
Observed band size: 55 kDa

Exposure time: 43 seconds;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750283) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:50,000 dilution was used for 1 hour at room temperature.

Fig2: This data was developed using HA750283, the same antibody clone in a different buffer formulation.
Western blot analysis of IRF3 on different lysates with Rabbit anti-IRF3 antibody (HA750283) at 1/5,000 dilution.

Lane 1: Hela-si NT cell lysate
Lane 2: Hela-si IRF3 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 47 kDa
Observed band size: 55 kDa

Exposure time: 3 minutes;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1612-14, 1/5,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

Fig3: This data was developed using HA750283, the same antibody clone in a different buffer formulation.
Western blot analysis of IRF3 on different lysates with Rabbit anti-IRF3 antibody (HA750283) at 1/5,000 dilution.

Lane 1: RAW264.7 cell lysate (20 µg/Lane)
Lane 2: Mouse spleen tissue lysate (40 µg/Lane)
Lane 3: Mouse liver tissue lysate (40 µg/Lane)

Predicted band size: 47 kDa
Observed band size: 55 kDa

Exposure time: 10 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750283) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig4: This data was developed using HA750283, the same antibody clone in a different buffer formulation.
Immunocytochemistry analysis of Jurkat cells labeling IRF3 with Rabbit anti-IRF3 antibody (HA750283) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-IRF3 antibody (HA750283) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

	Fig5: This data was developed using HA750283, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-IRF3 antibody (HA750283) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA750283) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig6: This data was developed using HA750283, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-IRF3 antibody (HA750283) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA750283) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig7: This data was developed using HA750283, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Rabbit anti-IRF3 antibody (HA750283) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA750283) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig8: This data was developed using HA750283, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-IRF3 antibody (HA750283) at 1/2,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA750283) at 1/2,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig9: This data was developed using HA750283, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded rat colon tissue with Rabbit anti-IRF3 antibody (HA750283) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA750283) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig10: This data was developed using HA750283, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded rat heart tissue with Rabbit anti-IRF3 antibody (HA750283) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA750283) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

	Fig11: This data was developed using HA750283, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-IRF3 antibody (HA750283) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA750283) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
	Fig12: This data was developed using HA750283, the same antibody clone in a different buffer formulation. Flow cytometric analysis of Jurkat cells labeling IRF3. Cells were fixed and permeabilized. Then stained with the primary antibody (HA750283, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
	Fig13: This data was developed using HA750283, the same antibody clone in a different buffer formulation. IRF3 was immunoprecipitated from 0.2 mg Jurkat cell lysate with HA750283 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA750283 at 1/5,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: Jurkat cell lysate (input) Lane 2: HA750283 IP in Jurkat cell lysate Lane 3: Rabbit IgG instead of HA750283 in Jurkat cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 9 seconds; ECL: K1801

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.