- 44-308G - Invitrogen Antibodies OPRM1 antibody

Submitted references Effects of electroacupuncture on pain sensation in a rat model of hyperalgesia with nicotine dependence.

Wang SJ, Zhang YP, Candiotti KA
Neural regeneration research 2022 Apr;17(4):905-910

Inhibition of mu-opioid receptor suppresses proliferation of hepatocellular carcinoma cells via CD147-p53-MAPK cascade signaling pathway.

Zhang JJ, Song CG, Dai JM, Zhang XQ, Lin P, Li L, Yang XM, Chen ZN
American journal of translational research 2021;13(5):3967-3986

Changes in midbrain pain receptor expression, gait and behavioral sensitivity in a rat model of radiculopathy.

Hwang PY, Allen KD, Shamji MF, Jing L, Mata BA, Gabr MA, Huebner JL, Kraus VB, Richardson WJ, Setton LA
The open orthopaedics journal 2012;6:383-91

Familial hemiplegic migraine type 1 mutations W1684R and V1696I alter G protein-mediated regulation of Ca(V)2.1 voltage-gated calcium channels.

Garza-López E, Sandoval A, González-Ramírez R, Gandini MA, Van den Maagdenberg A, De Waard M, Felix R
Biochimica et biophysica acta 2012 Aug;1822(8):1238-46

Quantitative Detection of µ Opioid Receptor: Western Blot Analyses Using µ Opioid Receptor Knockout Mice.

Kasai S, Yamamoto H, Kamegaya E, Uhl GR, Sora I, Watanabe M, Ikeda K
Current neuropharmacology 2011 Mar;9(1):219-22

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot. Extracts of rat brain lysates were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature and incubated with the µ-opioid receptor antibody for two hours at room temperature in a 1% BSA-TBST buffer. After washing, the membrane was incubated with goat F (ab’)2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and signals were detected using the Pierce SuperSignal™ method

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot. Extracts of rat brain lysates were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% BSA-TBST buffer for one hour at room temperature and incubated with the µ-opioid receptor antibody for two hours at room temperature in a 1% BSA-TBST buffer. After washing, the membrane was incubated with goat F (ab’)2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and signals were detected using the Pierce SuperSignal™ method

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of Neuro-2a (Lane 1), U-87 MG (Lane 2), IMR-32 (Lane 3), SK-N-AS (Lane 4), U-2 OS (Lane 5) and tissue extracts of Mouse Cerebellum (Lane 6) and Rat Brain (Lane 7). The blot was probed with Anti-Opioid Receptor Rabbit Polyclonal Antibody (Product # 44-308G, 1:250 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/mL, 1:2500 dilution). Opioid Receptor has multiple isoforms ranging from 20 to 55 kDa, which appears as a ladder form across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE and reg; 10% Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescence analysis of OPIOID RECEPTOR was performed using 70% confluent log phase Neuro-2A cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with OPRM1 Rabbit Polyclonal Antibody (Product # 44-308G) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing membranous localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 4 Electroacupuncture effects on MOR, beta-EP, GAD67, and GAD65 protein expression in the spinal cord of nicotine-induced hyperalgesic rats . (A-D) Western blot analysis of MOR (A), beta-EP (B), GAD67 (C), and GAD65 (D) in the spinal cord of rats in the three groups. Data (mean +- SEM) are presented as relative values to control levels (CTR: n = 6, NIC: n = 6, NIC + EA: n = 6). * P < 0.05, ** P < 0.01, vs . CTR group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs . NIC group (one-way analysis of variance followed by Newman-Keuls multiple comparison test). CTR: Age-matched control group; NIC: nicotine-exposed group; NIC + EA: nicotine-exposed/electroacupuncture-treated group. GAD65: glutamic acid decarboxylase 65; GAD67: glutamic acid decarboxylase 67; MOR: mu-opioid receptor; beta-EP: beta-endorphin.

Supportive validation

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Experimental details: Fig. (5) Mu-Opioid (u-OR) Receptor Immunostaining: PAG: (left) Representative images of u-OR immunohistochemical images for sham( A ) and NP-treated ( B ) 4 weeks after surgery (dark spots represent positive staining; arrows indicate representative positive staining; scalebar = 100 um). Increased expression was observed in both sham (* p

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. (3) Western blot analyses with N38 and AB5511 antibodies for MOP detection in a protein dose-dependent manner. The P2 fractions were isolated from the cortex of adult male C57BL/6J mice. 0.3, 1, 3, 10, and 30 ug of proteins were subjected to SDS-PAGE. Protein blots were reacted with N38 (1:100) and AB5511 (1:5,000) antibodies, followed by reaction with secondary antibody and visualization ( A ). The optical density of migrating bands indicated by arrows around 60-67 kDa was quantified using Image J software ( B ).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. (1) Western blot analyses with three anti-MOP antibodies produced against peptides from different MOP regions. Anti-MOP rabbit polyclonal antibodies (N38, 44-308G, and AB5511) were produced against the N -terminus, internal region, and C -terminus of MOP, respectively. Total cortex homogenate (T) from adult male C57BL/6J mice was fractionated to nuclear fraction (P1), cytoplasmic membrane fraction (P2), and cytoplasm fraction (S2) by two-step centrifugation at 800 x g and subsequently 22,000 x g . 10 ug of each fraction was subjected to SDS-PAGE. Protein blots were incubated with anti-MOP antibodies at appropriate dilution (N38, 1:400; 44-308G, 1:5,000; AB5511, 1:20,000), followed by reaction with secondary antibody and visualization.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig. (2) Western blot analyses at the P2 fraction derived from wildtype and Oprm1 knockout mice. The P2 fractions were isolated from the cortex of wildtype (WT) and Oprm1 knockout (KO) adult male mice. 10 ug of the P2 fraction from each genotype was subjected to SDS-PAGE. Protein blots were reacted with anti-MOP antibodies at appropriate dilution (N38, 1:400; 44-308G, 1:5,000; AB5511, 1:20,000), followed by reaction with secondary antibody and visualization.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 4 Electroacupuncture effects on MOR, beta-EP, GAD67, and GAD65 protein expression in the spinal cord of nicotine-induced hyperalgesic rats . (A-D) Western blot analysis of MOR (A), beta-EP (B), GAD67 (C), and GAD65 (D) in the spinal cord of rats in the three groups. Data (mean +- SEM) are presented as relative values to control levels (CTR: n = 6, NIC: n = 6, NIC + EA: n = 6). * P < 0.05, ** P < 0.01, vs . CTR group; # P < 0.05, ## P < 0.01, ### P < 0.001, vs . NIC group (one-way analysis of variance followed by Newman-Keuls multiple comparison test). CTR: Age-matched control group; NIC: nicotine-exposed group; NIC + EA: nicotine-exposed/electroacupuncture-treated group. GAD65: glutamic acid decarboxylase 65; GAD67: glutamic acid decarboxylase 67; MOR: mu-opioid receptor; beta-EP: beta-endorphin.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 5 Electroacupuncture effects on MOR, beta-EP, GAD67, and GAD65 protein expression in the PAG of nicotine-induced hyperalgesic rats . (A-D) Western blot analysis of MOR (A), beta-EP (B), GAD67 (C), and GAD65 (D) in the PAG of rats in the three groups. Data (mean +- SEM) are presented as relative values to control levels (CTR: n = 6, NIC: n = 6, NIC + EA: n = 6). * P < 0.05, ** P < 0.01, vs . CTR group; # P < 0.05, ## P < 0.01, vs . NIC group (one-way analysis of variance followed by Newman-Keuls multiple comparison test). CTR: Age-matched control group; NIC: nicotine-exposed group; NIC + EA: nicotine-exposed/electroacupuncture-treated group. GAD65: glutamic acid decarboxylase 65; GAD67: glutamic acid decarboxylase 67; MOR: mu-opioid receptor; PAG: periaqueductal gray; beta-EP: beta-endorphin.

44-308G

Antibody data