ET1601-22

p75 NGF Receptor Recombinant Rabbit Monoclonal Antibody [SA39-02]

cat.: ET1601-22

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality:	Monoclonal
Clone number:	SA39-02

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. 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 p75 NGF Receptor aa 345-394 / 427.
Positive control:	Neuro-2a cell lysates, PC-12 cell lysates, Hela, N2A, rat uterus tissue, human endometrium tissue, human tonsil tissue, mouse uterus, PC-12.
Subcellular location:	Membrane
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P IP FC	1:2,000-1:5,000 1:50-1:100 1:50-1:100 1:200-1:1,000 Use at an assay dependent concentration. 1:200
Uniprot #:	SwissProt: P08138 Human \| Q9Z0W1 Mouse \| P07174 Rat
Alternative names:	CD271 CD271 antigen Gp80 LNGFR Gp80-LNGFR Low affinity nerve growth factor receptor Low affinity neurotrophin receptor p75NTR Low-affinity nerve growth factor receptor Nerve growth factor receptor Nerve growth factor receptor TNFR superfamily member 16 NGF receptor Ngfr p75 ICD p75 Neurotrophin receptor p75 NTR p75(NTR) p75NTR TNFR Superfamily Member 16 TNFRSF16 TNR16_HUMAN Tumor necrosis factor receptor superfamily member 16

Images

Fig1: Western blot analysis of p75 NGF Receptor on Neuro-2a cell lysates with Rabbit anti-p75 NGF Receptor antibody (ET1601-22) at 1/2,000 dilution.

Lysates/proteins at 15 µg/Lane.

Predicted band size: 45 kDa
Observed band size: 70 kDa

Exposure time: 3 minutes;

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 (ET1601-22) at 1/2,000 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 p75 NGF Receptor on PC-12 cell lysates with Rabbit anti-p75 NGF Receptor antibody (ET1601-22) at 1/5,000 dilution.

Lysates/proteins at 15 µg/Lane.

Predicted band size: 45 kDa
Observed band size: 70 kDa

Exposure time: 1 minute;

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 (ET1601-22) 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.

	Fig3: Western blot analysis of p75 NGF Receptor on different lysates with Rabbit anti-p75 NGF Receptor antibody (ET1601-22) at 1/2,000 dilution. Lane 1: PC-12-si NT cell lysate Lane 2: PC-12-si p75 NGF Receptor cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 45 kDa Observed band size: 70 kDa Exposure time: 2 minutes 6 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 (ET1601-22) at 1/2,000 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: Immunocytochemistry analysis of Hela cells labeling p75 NGF Receptor with Rabbit anti-p75 NGF Receptor antibody (ET1601-22) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-p75 NGF Receptor antibody (ET1601-22) at 1/100 dilution in 2% negative goat serum 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.
	Fig5: ICC staining of p75 NGF Receptor in N2A cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1601-22, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).

	Fig6: Immunohistochemical analysis of paraffin-embedded rat uterus tissue with Rabbit anti-p75 NGF Receptor antibody (ET1601-22) 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 (ET1601-22) 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 human endometrium tissue with Rabbit anti-p75 NGF Receptor antibody (ET1601-22) at 1/500 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 (ET1601-22) at 1/500 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 human tonsil tissue with Rabbit anti-p75 NGF Receptor antibody (ET1601-22) 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 (ET1601-22) 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.

	Fig9: Immunohistochemical analysis of paraffin-embedded mouse uterus tissue using anti-p75 NGF Receptor antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) 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 (ET1601-22, 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.
	Fig10: Flow cytometric analysis of PC-12 cells labeling p75 NGF Receptor. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1601-22, 1:200) (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: Western blot analysis of p75 NGF Receptor on SH-SY5Y cell lysates with Rabbit anti-p75 NGF Receptor antibody (ET1601-22) at 1/2,000 dilution. Lysates/proteins at 20µg/Lane. Predicted band size: 45 kDa Observed band size: 70 kDa Exposure time: 1 minutes 59 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 (ET1601-22) at 1/2,000 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.

Fig12: Immunocytochemistry analysis of Neuro-2a cells labeling p75 NGF Receptor with Rabbit anti-p75 NGF Receptor antibody (ET1601-22) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-p75 NGF Receptor antibody (ET1601-22) at 1/100 dilution in 2% negative goat serum 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.

Fig13: Immunocytochemistry analysis of PC-12 cells labeling p75 NGF Receptor with Rabbit anti-p75 NGF Receptor antibody (ET1601-22) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-p75 NGF Receptor antibody (ET1601-22) at 1/100 dilution in 2% negative goat serum 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.

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