HA721813

NSDHL Recombinant Rabbit Monoclonal Antibody [PSH02-36]

cat.: HA721813

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell, FC
Clonality:	Monoclonal
Clone number:	PSH02-36

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 42 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human NSDHL aa 1-300 / 373 (Q15738).
Positive control:	HeLa cell lysate, HepG2 cell lysate, A431 cell lysate, HEK-293 cell lysate, AGS cell lysate, MCF7 cell lysate, MDA-MB-231 cell lysate, NIH/3T3 cell lysate, bEnd.3 cell lysate, PC-12 cell lysate, C6 cell lysate, human liver tissue lysate, mouse liver tissue lysate, rat liver tissue lysate, human colon cancer tissue, human liver cancer tissue, human liver tissue, human stomach cancer tissue, mouse liver tissue, rat liver tissue, HepG2, NIH/3T3.
Subcellular location:	Endoplasmic reticulum membrane, Lipid droplet
Recommended Dilutions: WB IHC-P IF-Cell FC	1:1,000 1:200-1:1,000 1:100 1:100-1:1,000
Uniprot #:	SwissProt: Q15738 Human \| Q9R1J0 Mouse \| Q5PPL3 Rat
Alternative names:	decarboxylating H105E3 H105e3 protein NAD(P) dependent steroid dehydrogenase like NSDHL NSDHL_HUMAN Protein H105e3 SDR31E1 Short chain dehydrogenase/reductase family 31E member 1 Sterol 4 alpha carboxylate 3 dehydrogenase decarboxylating Sterol-4-alpha-carboxylate 3-dehydrogenase XAP104

Images

Fig1: Western blot analysis of NSDHL on different lysates with Rabbit anti-NSDHL antibody (HA721813) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HepG2 cell lysate
Lane 3: A431 cell lysate
Lane 4: HEK-293 cell lysate
Lane 5: AGS cell lysate
Lane 6: MCF7 cell lysate
Lane 7: MDA-MB-231 cell lysate
Lane 8: NIH/3T3 cell lysate
Lane 9: bEnd.3 cell lysate
Lane 10: PC-12 cell lysate
Lane 11: C6 cell lysate
Lane 12: Human liver tissue lysate
Lane 13: Mouse liver tissue lysate
Lane 14: Rat liver tissue lysate

Lysates/proteins at 30 µg/Lane.

Predicted band size: 42 kDa
Observed band size: 40 kDa

Exposure time: 42 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 (HA721813) 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: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-NSDHL antibody (HA721813) at 1/1,000 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 (HA721813) 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.

	Fig3: Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-NSDHL antibody (HA721813) at 1/1,000 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 (HA721813) 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 liver tissue with Rabbit anti-NSDHL antibody (HA721813) at 1/1,000 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 (HA721813) 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 human stomach cancer tissue with Rabbit anti-NSDHL antibody (HA721813) at 1/200 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 (HA721813) 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.

	Fig6: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-NSDHL antibody (HA721813) at 1/1,000 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 (HA721813) 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: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-NSDHL antibody (HA721813) at 1/1,000 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 (HA721813) 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.
	Fig8: Immunocytochemistry analysis of HepG2 cells labeling NSDHL with Rabbit anti-NSDHL antibody (HA721813) 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-NSDHL antibody (HA721813) 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 NSDHL with Rabbit anti-NSDHL antibody (HA721813) 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-NSDHL antibody (HA721813) 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: Flow cytometric analysis of HepG2 cells labeling NSDHL. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721813, 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 NIH/3T3 cells labeling NSDHL. Cells were fixed and permeabilized. Then stained with the primary antibody (HA721813, 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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