Anti-NGFR/p75 neurotrophin receptor (p75NTR) Antibody (8J2)

Our Anti-NGFR/p75 neurotrophin receptor (p75NTR) mouse monoclonal primary antibody detects human, mouse, and rat NGFR/p75 neurotrophin receptor (p75NTR), and is IgG. It is validated for use in FC, ICC, IHC-Frozen, Immunopanning, IP, WB.

Left: Analysis of p75NTR expression in human and rodent RIPA cell lysates by Western Blotting. Mouse antibody to p75NTR clone 8J2 (M-1818-100) and clone MLR2 (M-009-100) detect p75NTR-IR at ~50-60 kDa in mouse p75NTR-transfected cells (A), but not in non-transfected control cells (B). p75NTR-IR is also observed in human SH-SY5Y (C) and rat C6 (D) cell lysates. Dependent on cell lysate, dimers or trimers (~150 kDa) of p75NTR are detected (Anastasia et al., 2015). 50 ug of protein were loaded per lane. WB Method: SDS-PAGE: 4-12%, non-reducing conditions; Transfer: Tris-Glycine buffer; Membrane: nitrocellulose (0.45 um); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: 1 µg/mL, overnight at 4°C; Secondary antibody: anti-mouse-HRP (1/6000) 1 hour at RT; Detection: Chemiluminiscence. Right: Immunoprecipitation of p75NTR from rat C6 cell lysate and detection by Western Blotting. Mouse antibody to p75NTR clone 8J2 (M-1818-100) and clone MLR2 (M-009-100) precipitate bands at the expected molecular weight of ~50-60 kDa for p75NTR (A). No p75NTR-IR is observed in remaining supernatant (B) or control (Protein G only, C). IP Method: C6 lysates were prepared in non-denaturing lysis buffer and pre-cleared with Protein G agarose beads (40 uL bead slurry per 1 mg total protein). Cleared lysate was then incubated with either M-1818-100 or M-009-100 (10 ug antibody per 200 ug total protein), and immune complexes bound by Protein G agarose (10 uL). Precipitated p75NTR was eluted off the beads and antibody by heating and addition of SDS-PAGE sample buffer. WB Method: as above, with primary goat anti-p75NTR (1 µg/mL) antibody and secondary anti-sheep-HRP (1/6000) antibody. Detection: Chemiluminiscence.
Binding analysis of M-1818-100 (8J2) and M-009-100 (MLR2) antibodies by 1-site ELISA on human and mouse p75NTR protein. Clone 8J2 detects both, human and mouse p75NTR proteins with higher affinity than clone MLR2.
Analysis of membrane p75NTR expression in rat C6 cells by Immunocytochemistry. Non-permeabilized cells were stained with mouse antibodies to p75NTR clones 8J2 (M-1818-100) and MLR2 (M-009-100) either on ice without fixation (top panel), or after fixation with 4% formaldehyde (bottom panel). Both antibodies reveal punctuate membrane staining of p75NTR receptor. This staining is specific, because negative mouse control antibody clone X63 (M-1249-100) does not show immunoreactivity. Method: Primary antibodies: 2 µg/mL, 1 hour incubation; Blocking: 10% normal horse serum, 30 minutes; Fixation: 4% formaldehyde, 10 minutes, either before application of primary antibody (bottom panel) or after application of primary antibody (upper panel); Secondary antibody: donkey anti-mouse-CF488A (2 µg/mL), 1 hour incubation. Cell nuclei were stained with Hoechst dye (blue), 1:1,000, 20 minutes at room temperature. Magnification: 100x.
p75NTR staining (green) in rat trigeminal ganglion cells by Immunocytochemistry. Primary antibody dilution: 1:100. Image courtesy of QBM Cell Science.
Left: Analysis of p75NTR expression in human and rodent RIPA cell lysates by Western Blotting. Mouse antibody to p75NTR clone 8J2 (M-1818-100) and clone MLR2 (M-009-100) detect p75NTR-IR at ~50-60 kDa in mouse p75NTR-transfected cells (A), but not in non-transfected control cells (B). p75NTR-IR is also observed in human SH-SY5Y (C) and rat C6 (D) cell lysates. Dependent on cell lysate, dimers or trimers (~150 kDa) of p75NTR are detected (Anastasia et al., 2015). 50 ug of protein were loaded per lane. WB Method: SDS-PAGE: 4-12%, non-reducing conditions; Transfer: Tris-Glycine buffer; Membrane: nitrocellulose (0.45 um); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: 1 µg/mL, overnight at 4°C; Secondary antibody: anti-mouse-HRP (1/6000) 1 hour at RT; Detection: Chemiluminiscence. Right: Immunoprecipitation of p75NTR from rat C6 cell lysate and detection by Western Blotting. Mouse antibody to p75NTR clone 8J2 (M-1818-100) and clone MLR2 (M-009-100) precipitate bands at the expected molecular weight of ~50-60 kDa for p75NTR (A). No p75NTR-IR is observed in remaining supernatant (B) or control (Protein G only, C). IP Method: C6 lysates were prepared in non-denaturing lysis buffer and pre-cleared with Protein G agarose beads (40 uL bead slurry per 1 mg total protein). Cleared lysate was then incubated with either M-1818-100 or M-009-100 (10 ug antibody per 200 ug total protein), and immune complexes bound by Protein G agarose (10 uL). Precipitated p75NTR was eluted off the beads and antibody by heating and addition of SDS-PAGE sample buffer. WB Method: as above, with primary goat anti-p75NTR (1 µg/mL) antibody and secondary anti-sheep-HRP (1/6000) antibody. Detection: Chemiluminiscence.
Left: Analysis of p75NTR expression in human and rodent RIPA cell lysates by Western Blotting. Mouse antibody to p75NTR clone 8J2 (M-1818-100) and clone MLR2 (M-009-100) detect p75NTR-IR at ~50-60 kDa in mouse p75NTR-transfected cells (A), but not in non-transfected control cells (B). p75NTR-IR is also observed in human SH-SY5Y (C) and rat C6 (D) cell lysates. Dependent on cell lysate, dimers or trimers (~150 kDa) of p75NTR are detected (Anastasia et al., 2015). 50 ug of protein were loaded per lane. WB Method: SDS-PAGE: 4-12%, non-reducing conditions; Transfer: Tris-Glycine buffer; Membrane: nitrocellulose (0.45 um); Blocking: 5% skim milk in TBST, 1 hour at RT; Primary antibody: 1 µg/mL, overnight at 4°C; Secondary antibody: anti-mouse-HRP (1/6000) 1 hour at RT; Detection: Chemiluminiscence. Right: Immunoprecipitation of p75NTR from rat C6 cell lysate and detection by Western Blotting. Mouse antibody to p75NTR clone 8J2 (M-1818-100) and clone MLR2 (M-009-100) precipitate bands at the expected molecular weight of ~50-60 kDa for p75NTR (A). No p75NTR-IR is observed in remaining supernatant (B) or control (Protein G only, C). IP Method: C6 lysates were prepared in non-denaturing lysis buffer and pre-cleared with Protein G agarose beads (40 uL bead slurry per 1 mg total protein). Cleared lysate was then incubated with either M-1818-100 or M-009-100 (10 ug antibody per 200 ug total protein), and immune complexes bound by Protein G agarose (10 uL). Precipitated p75NTR was eluted off the beads and antibody by heating and addition of SDS-PAGE sample buffer. WB Method: as above, with primary goat anti-p75NTR (1 µg/mL) antibody and secondary anti-sheep-HRP (1/6000) antibody. Detection: Chemiluminiscence.
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Human, Mouse, Rat
Flow, ICC, IHC, IP, WB
Mouse

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Data Sheet

SKU: M-1818-100

Volume/Size: 100 µg
Price:
$337.00

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