HA722394

NAP1L1 Recombinant Rabbit Monoclonal Antibody [JE34-44]

cat.: HA722394

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC, IF-Tissue
Clonality:	Monoclonal
Clone number:	JE34-44

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: 45 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human NAP1L1 aa 1-100.
Positive control:	HeLa cell lysate, Jurkat cell lysate, HepG2 cell lysate, 293T cell lysate, A431 cell lysate, NIH/3T3 cell lysate, C6 cell lysate, HeLa, NIH/3T3, C6, human testis tissue, mouse testis tissue, rat testis tissue.
Subcellular location:	Nucleus, Melanosome, Cytoplasm.
Recommended Dilutions: WB IF-Cell IHC-P FC IF-Tissue	1:1,000 1:100 1:5,000 1:1,000 1:200-1:1,000
Uniprot #:	SwissProt: P55209 Human \| P28656 Mouse \| Q9Z2G8 Rat
Alternative names:	FLJ16112 hNRP HSP22 like protein interacting protein MGC23410 MGC8688 NAP 1 NAP 1 related protein NAP 1L NAP-1-related protein NAP1 NAP1 L1 NAP1 related protein NAP1L Nap1l1 NAP1L1 protein NP1L1_HUMAN NRP Nucleosome assembly protein 1 like 1 Nucleosome assembly protein 1-like 1 Nucleosome assembly protein I-related protein

Images

Fig1: Western blot analysis of NAP1L1 on different lysates with Rabbit anti-NAP1L1 antibody (HA722394) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: Jurkat cell lysate
Lane 3: HepG2 cell lysate
Lane 4: 293T cell lysate
Lane 5: A431 cell lysate
Lane 6: NIH/3T3 cell lysate
Lane 7: C6 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 45 kDa
Observed band size: 50/55 kDa

Exposure time: 6 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 (HA722394) 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: Immunocytochemistry analysis of HeLa cells labeling NAP1L1 with Rabbit anti-NAP1L1 antibody (HA722394) 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-NAP1L1 antibody (HA722394) 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.

	Fig3: Immunocytochemistry analysis of NIH/3T3 cells labeling NAP1L1 with Rabbit anti-NAP1L1 antibody (HA722394) 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-NAP1L1 antibody (HA722394) 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.
	Fig4: Immunocytochemistry analysis of C6 cells labeling NAP1L1 with Rabbit anti-NAP1L1 antibody (HA722394) 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-NAP1L1 antibody (HA722394) 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: Flow cytometric analysis of HeLa cells labeling NAP1L1. Cells were fixed and permeabilized. Then stained with the primary antibody (HA722394, 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).

	Fig6: Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-NAP1L1 antibody (HA722394) at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722394) at 1/5,000 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: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-NAP1L1 antibody (HA722394) at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722394) at 1/5,000 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.
	Fig8: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-NAP1L1 antibody (HA722394) at 1/5,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 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 (HA722394) at 1/5,000 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.

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