HA720027

RCL Recombinant Rabbit Monoclonal Antibody [JE59-29]

cat.: HA720027

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JE59-29

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: 19 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human RCL aa 75-174/174.
Positive control:	PC-12 cell lysates, K562 cell lysate, Jurkat cell lysate, 293T cell lysate, HepG2 cell lysate, human liver tissue, human kidney tissue, HepG2, C2C12, PC-12.
Subcellular location:	Nucleus, Cytoplasm.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:1,000-1:5,000 1:100-1:200 1:50-1:200 1:1,000
Uniprot #:	SwissProt: O43598 Human \| Q80VJ3 Mouse \| O35820 Rat
Alternative names:	2' deoxynucleoside 5' phosphate N hydrolase 1 BC048355 c Myc responsive c Myc responsive protein Rcl c-Myc-responsive protein Rcl C6orf108 C76683 Chromosome 6 open reading frame 108 Deoxyribonucleoside 5' monophosphate N glycosidase Deoxyribonucleoside 5''-monophosphate N-glycosidase dJ330M21.3 DNPH1 MGC54855 Putative c Myc responsive Rcl RCL_HUMAN RP3 330M21.3

Images

Fig1: Western blot analysis of RCL on different lysates with Rabbit anti-RCL antibody (HA720027) at 1/1,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-RCL KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 19 kDa
Observed band size: 19 kDa

Exposure time: 180 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 (HA720027) at 1/1,000 dilution was used in K1803 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 RCL on PC-12 cell lysates with Rabbit anti-RCL antibody (HA720027) at 1/5,000 dilution.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 19 kDa
Observed band size: 19 kDa

Exposure time: 13 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 (HA720027) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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: Western blot analysis of RCL on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (HA720027, 1/1,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature. Positive control: Lane 1: K562 cell lysate Lane 2: Jurkat cell lysate Lane 3: 293T cell lysate Lane 4: HepG2 cell lysate
	Fig4: Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-RCL antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA720027, 1/200) for 30 minutes 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 human kidney tissue using anti-RCL antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (HA720027, 1/200) for 30 minutes 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: Immunocytochemistry analysis of HepG2 cells labeling RCL with Rabbit anti-RCL antibody (HA720027) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-RCL antibody (HA720027) 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 (HA601187, 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.

Fig7: Immunocytochemistry analysis of C2C12 cells labeling RCL with Rabbit anti-RCL antibody (HA720027) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-RCL antibody (HA720027) 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 (HA601187, 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.

	Fig8: Immunocytochemistry analysis of PC-12 cells labeling RCL with Rabbit anti-RCL antibody (HA720027) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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-RCL antibody (HA720027) 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 (HA601187, 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: Flow cytometric analysis of HepG2 cells labeling RCL. Cells were fixed and permeabilized. Then stained with the primary antibody (HA720027, 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).
	Fig10: Flow cytometric analysis of C2C12 cells labeling RCL. Cells were fixed and permeabilized. Then stained with the primary antibody (HA720027, 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).

Fig11: Flow cytometric analysis of PC-12 cells labeling RCL.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA720027, 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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