HA722552

CUG-BP1 Recombinant Rabbit Monoclonal Antibody [JE40-85]

cat.: HA722552

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell, FC, IP
Clonality:	Monoclonal
Clone number:	JE40-85

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 52 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within
Positive control:	HeLa cell lysate, 293T cell lysate, SH-SY5Y cell lysate, NCI-H226 cell lysate, COS-1 cell lysate, Neuro-2a cell lysate, NIH/3T3 cell lysate, C6 cell lysate, L6 cell lysate, PC-12 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, human kidney tissue, human brain tissue, mouse brain tissue, rat brain tissue, HeLa, NIH/3T3, C6.
Subcellular location:	Nucleus, Cytoplasm.
Recommended Dilutions: WB IHC-P IF-Cell FC IP	1:1,000 1:1,000 1:100 1:1,000 1-2μg/sample
Uniprot #:	SwissProt: Q92879 Human \| P28659 Mouse \| Q4QQT3 Rat
Alternative names:	50 kDa Nuclear polyadenylated RNA binding protein 50 kDa nuclear polyadenylated RNA-binding protein Bruno like 2 bruno like protein 2 Bruno-like protein 2 BRUNOL 2 BRUNOL2 CELF 1 CELF-1 celf1 CELF1 CUGBP, Elav like family member 1 CELF1_HUMAN CUG BP and ETR 3 like factor 1 CUG BP CUG BP1 CUG RNA binding protein CUG triplet repeat RNA binding protein 1 CUG triplet repeat RNA-binding protein 1 CUG-BP CUG-BP- and ETR-3-like factor 1 CUG-BP1 CUGBP 1 CUGBP and ETR3 like factor 1 CUGBP CUGBP Elav like family member 1 CUGBP Elav-like family member 1 CUGBP1 Cytidine uridine guanosine binding protein 1 Deadenylation factor CUG BP Deadenylation factor CUG-BP Deadenylation factor CUGBP EDEN BP EDEN BP homolog EDEN-BP EDEN-BP homolog embryo deadenylation element binding protein embryo deadenylation element binding protein homolog Embryo deadenylation element-binding protein homolog hNab 50 hNab50 NAB 50 NAB50 NAPOR......

Images

Fig1: Western blot analysis of CUG-BP1 on different lysates with Rabbit anti-CUG-BP1 antibody (HA722552) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: 293T cell lysate
Lane 3: SH-SY5Y cell lysate
Lane 4: NCI-H226 cell lysate
Lane 5: COS-1 cell lysate
Lane 6: Neuro-2a cell lysate
Lane 7: NIH/3T3 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 52 kDa
Observed band size: 52 kDa

Exposure time: 1 minute 34 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 (HA722552) at 1/1,000 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: Western blot analysis of CUG-BP1 on different lysates with Rabbit anti-CUG-BP1 antibody (HA722552) at 1/1,000 dilution.

Lane 1: C6 cell lysate (20 µg/Lane)
Lane 2: L6 cell lysate (20 µg/Lane)
Lane 3: PC-12 cell lysate (20 µg/Lane)
Lane 4: Mouse brain tissue lysate (30 µg/Lane)
Lane 5: Rat brain tissue lysate (30 µg/Lane)

Predicted band size: 52 kDa
Observed band size: 52 kDa

Exposure time: 2 minutes; 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 (HA722552) at 1/1,000 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.

	Fig3: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-CUG-BP1 antibody (HA722552) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (HA722552) at 1/1,000 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-CUG-BP1 antibody (HA722552) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (HA722552) at 1/1,000 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-CUG-BP1 antibody (HA722552) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (HA722552) at 1/1,000 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: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-CUG-BP1 antibody (HA722552) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) 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 (HA722552) at 1/1,000 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: CUG-BP1 was immunoprecipitated from 0.2 mg HeLa cell lysate with HA722552 at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using HA722552 at 1/1,000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody (NBI01H) at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input) Lane 2: HA722552 IP in HeLa cell lysate Lane 3: Rabbit IgG instead of HA722552 in HeLa cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 1 minute 5 seconds; ECL: K1802
	Fig8: Immunocytochemistry analysis of HeLa cells labeling CUG-BP1 with Rabbit anti-CUG-BP1 antibody (HA722552) 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-CUG-BP1 antibody (HA722552) 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.

	Fig9: Immunocytochemistry analysis of NIH/3T3 cells labeling CUG-BP1 with Rabbit anti-CUG-BP1 antibody (HA722552) 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-CUG-BP1 antibody (HA722552) 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.
	Fig10: Immunocytochemistry analysis of C6 cells labeling CUG-BP1 with Rabbit anti-CUG-BP1 antibody (HA722552) 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-CUG-BP1 antibody (HA722552) 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.
	Fig11: Flow cytometric analysis of HeLa cells labeling CUG-BP1. Cells were fixed and permeabilized. Then stained with the primary antibody (HA722552, 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).

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