rabbit anti-cb2 antibody  (Cayman Chemical)

Journal: Acta histochemica

Article Title: Immunohistochemical distribution of cannabinoid receptor type 1 (CB1) and type 2 (CB2) in the rat carotid body.

doi: 10.1016/j.acthis.2024.152205

Figure Lengend Snippet: Fig. 1. Immunohistochemistry for CB1 and CB2 in the carotid body. (A) Dot-like CB1 immunoreactivity is observed throughout the carotid body. Intense immu- noreactivity is shown in nerve fibers and/or endings (arrows). (B) A higher magnification view of the rectangle in panel A. Dot-like CB1 immunoreactivity is observed in the perinuclear cytoplasm of chemoreceptor cells (arrows). Intense immunoreactivity for CB1 is also present in nerve endings (arrowheads). (C) Dot-like CB2 immunoreactivity is noted throughout the carotid body. (D) A higher magnification view (rectangle in panel C) shows small dots of CB2 immunoreactivity in the perinuclear region of chemoreceptor cells (arrow). (E-G) Double immunofluorescence for CB1 and CB2 shows CB1-immunoreactive (arrows) and CB2- immunoreactive dots (arrow heads) in the same chemoreceptor cell; however, they are not colocalized.

Article Snippet: CB2: A rabbit polyclonal anti-CB2 antibody (NB300–606, Novus Biologicals, Centennial, CO, U.S.A.) was raised from a fusion protein that contained the first 33 amino acid residues of human CB2.

Techniques: Immunohistochemistry, Immunofluorescence

Journal: Acta histochemica

Article Title: Immunohistochemical distribution of cannabinoid receptor type 1 (CB1) and type 2 (CB2) in the rat carotid body.

doi: 10.1016/j.acthis.2024.152205

Figure Lengend Snippet: Fig. 2. (A-C)Triple immunofluorescence for CB1 with TH and DBH. CB1-immunoreactive dot-like structures are observed in both TH- (arrows) and DBH- immunoreactive chemoreceptor cells (arrowheads). Dot-like immunoreactivity for CB1 localizes in both the perinuclear cytoplasm and outlines of the regions of immunoreactivity for TH or DBH. (D-F) Triple immunofluorescence for CB2 with TH and DBH. CB2 immunoreactivity is shown in the perinuclear cytoplasm of TH- (arrows) and DBH-immunoreactive chemoreceptor cells (arrowheads).

Article Snippet: CB2: A rabbit polyclonal anti-CB2 antibody (NB300–606, Novus Biologicals, Centennial, CO, U.S.A.) was raised from a fusion protein that contained the first 33 amino acid residues of human CB2.

Techniques: Immunofluorescence

Journal: Acta histochemica

Article Title: Immunohistochemical distribution of cannabinoid receptor type 1 (CB1) and type 2 (CB2) in the rat carotid body.

doi: 10.1016/j.acthis.2024.152205

Figure Lengend Snippet: Fig. 3. (A-C) Double immunofluorescence for CB1 with P2X3. Intense CB1 immunoreactivity is surrounded by P2X3 immunoreactivity in the sensory nerve endings around chemoreceptor cells (arrows). (D-F) Double immunofluorescence for CB1 with VGluT2. CB1 immunoreactivity colocalizes with VGluT2 immunoreactivity, and appears to be in close contact with chemoreceptor cells (arrows). (G-H) Double immunofluorescence for CB2 with P2X2. Weak CB2-immunoreactive dots localize within P2X2-immunoreactive sensory nerve endings (arrows).

Article Snippet: CB2: A rabbit polyclonal anti-CB2 antibody (NB300–606, Novus Biologicals, Centennial, CO, U.S.A.) was raised from a fusion protein that contained the first 33 amino acid residues of human CB2.

Techniques: Immunofluorescence

Journal: Frontiers in Pharmacology

Article Title: Electroacupuncture reduces chronic itch via cannabinoid CB1 receptors in the ventrolateral periaqueductal gray

doi: 10.3389/fphar.2022.931600

Figure Lengend Snippet: Effect of EA on the expression of CB1 and CB2 receptor protein levels in the vlPAG. (A) The expression of c-Fos in the control group, AEW and EA group. (B) Representative gel image (upper) and quantification (bottom) of the protein level of CB1 receptors in control, AEW, EA and sham EA groups. (C) Representative gel image (upper) and quantification (bottom) of the protein level of CB2 receptors in control, AEW, EA and sham EA groups. GAPDH (36 kDa) was used as a loading control. The protein band at 60 kDa corresponds to the CB1 receptors protein. The protein band at 40 kDa corresponds to the CB2 receptors protein. Data are expressed as means ± SEM ( n = 6 mice in each group). * p < 0.05.

Article Snippet: Then, the transferred blot was blocked in Tris buffered saline (TBST) containing 5% skim milk powder at room temperature for 1 h. The membrane was placed in the following primary antibodies at 4°C overnight: rabbit anti-CB1 monoclonal antibody (1:1000, Cell Signaling Technology), rabbit anti-CB2 polyclonal antibody (1:500, Abcam), and mouse anti-GAPDH (0.5 μg/ml, Cloud-Clone Corp).

Techniques: Expressing, Control

Journal: The Journal of Biological Chemistry

Article Title: Cannabinoid CB2 receptors are upregulated via bivalent histone modifications and control primary afferent input to the spinal cord in neuropathic pain

doi: 10.1016/j.jbc.2022.101999

Figure Lengend Snippet: Nerve injury causes a delayed and sustained increase in CB2 expression in the DRG. A – C , representative gel images and quantification of CB2 protein level in the rat DRG 5 ( A ), 10 ( B ), and 21 ( C ) days after sham or SNL surgery. D , representative gel images and quantification of CB2 protein level in the rat dorsal spinal cord 21 days after sham or SNL surgery. Data are shown as means ± SEM (n = 9 rats per group). GAPDH was used as endogenous loading control, and the mean value in the sham group was normalized to 1. ∗ p < 0.05, ∗∗ p < 0.01, compared with the sham group (Two-tailed Student’s t test). CB2, Type-2 cannabinoid receptors; DRG, dorsal root ganglion; SNL, spinal nerve ligation.

Article Snippet: The membrane was incubated with blocking solution (5% nonfat dry milk in Tris-buffered saline and 0.05% Tween 20 (TBST)) at 22 °C for 1 h and then incubated overnight at 4 °C with an anti-rabbit CB2 antibody (1:200 dilution, #101550, Cayman Chemicals) diluted with TBST containing 3% bovine serum albumin.

Techniques: Expressing, Control, Two Tailed Test, Ligation

Journal: The Journal of Biological Chemistry

Article Title: Cannabinoid CB2 receptors are upregulated via bivalent histone modifications and control primary afferent input to the spinal cord in neuropathic pain

doi: 10.1016/j.jbc.2022.101999

Figure Lengend Snippet: Nerve injury increases CB2 expression in DRG neurons. A , representative low- and high-magnification images of RNAscope in situ hybridization show colabeling of NeuN ( green ) and CB2 mRNA ( red ) in DRG tissue sections from sham and SNL rats 10 days after surgery. B – D , quantification of CB2 mRNA punctate signals in the DRG tissue section ( B ), the abundance of CB2 mRNA signals in DRG neurons ( C ), and amount of CB2 mRNA signals in small-, medium-, and large-diameter DRG neurons ( D ) in sham and SNL rats. Data are shown as means ± SEM (n = 9 images per group). ∗ p < 0.05, ∗∗∗ p < 0.001 compared with the sham group (Two-tailed Student’s t test). CB2, Type-2 cannabinoid receptors; DRG, dorsal root ganglion; SNL, spinal nerve ligation.

Article Snippet: The membrane was incubated with blocking solution (5% nonfat dry milk in Tris-buffered saline and 0.05% Tween 20 (TBST)) at 22 °C for 1 h and then incubated overnight at 4 °C with an anti-rabbit CB2 antibody (1:200 dilution, #101550, Cayman Chemicals) diluted with TBST containing 3% bovine serum albumin.

Techniques: Expressing, RNAscope, In Situ Hybridization, Two Tailed Test, Ligation

Journal: The Journal of Biological Chemistry

Article Title: Cannabinoid CB2 receptors are upregulated via bivalent histone modifications and control primary afferent input to the spinal cord in neuropathic pain

doi: 10.1016/j.jbc.2022.101999

Figure Lengend Snippet: CB2 stimulation preferentially inhibits glutamatergic input from primary afferents to spinal dorsal horn neurons increased by nerve injury. A and B , representative recording traces ( A ) and mean data ( B ) show the effect of bath application of 10, 20, and 50 μM JWH-133 on the amplitude of EPSCs of dorsal horn neurons evoked monosynaptically from dorsal root stimulation in four sham (n = 12 neurons) or five SNL (n = 13 neurons) rats 2 to 3 weeks after surgery. C and D , original recording traces ( C ) and mean data ( D ) show the effect of bath application of 10 to 50 μM JWH-133 on the paired-pulse ratio of dorsal horn neurons in four sham (n = 11 neurons) or five SNL (n = 13 neurons) rats 2 to 3 weeks after surgery. Data are shown as means ± SEM. In (B), two-way ANOVA showed that there was a significant main effect for drug treatment (F(4,48) = 19.04, p < 0.001) and a significant interaction between SNL and sham conditions (F(12,48) = 6.27, p < 0.001). In (D), two-way ANOVA showed that there was a significant main effect for drug treatment (F(4,48) = 6.65, p < 0.001) and a significant interaction between SNL and sham conditions (F(12,48) = 5.56, p < 0.001). # p < 0.05, ### p < 0.001, compared with the baseline in sham rats. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, compared with the baseline in SNL rats (two-way ANOVA followed by the Dunnett’s post hoc test). E and F , representative blotting images ( E ) and quantification ( F ) of the CB2 protein level in dorsal spinal cord synaptosomes in SNL and sham control rats 21 days after surgery (n = 10 rats per group). Data are shown as mean ± SEM. PSD95, a synaptic protein, was used as a loading control. ∗∗ p < 0.01 (Two-tailed Student’s t test). CB2, Type-2 cannabinoid receptors; EPSCs, excitatory postsynaptic currents; SNL, spinal nerve ligation.

Article Snippet: The membrane was incubated with blocking solution (5% nonfat dry milk in Tris-buffered saline and 0.05% Tween 20 (TBST)) at 22 °C for 1 h and then incubated overnight at 4 °C with an anti-rabbit CB2 antibody (1:200 dilution, #101550, Cayman Chemicals) diluted with TBST containing 3% bovine serum albumin.

Techniques: Control, Two Tailed Test, Ligation

Journal: The Journal of Biological Chemistry

Article Title: Cannabinoid CB2 receptors are upregulated via bivalent histone modifications and control primary afferent input to the spinal cord in neuropathic pain

doi: 10.1016/j.jbc.2022.101999

Figure Lengend Snippet: CB2 activation at the spinal cord level attenuates pain hypersensitivity induced by nerve injury. A – C , time course of the effect of intrathecal injection of 10 to 100 μg JWH-133 on the paw withdrawal thresholds tested using von Frey filaments (A; 50 μg: p < 0.001, F(1.88,13.14) = 26.83; 100 μg: p < 0.001, F(1.26,8.84) = 22.82), a pressure stimulus (B; 50 μg: p < 0.001, F(2.52, 17.61) = 19.68; 100 μg: p < 0.001, F(2.65,18.54) = 26.30), and a heat stimulus (C; 50 μg: p = 0.0017, F(2.90,20.30) = 7.40; 100 μg: p < 0.001, F(2.97,20.81) = 10.95) in rats 2 to 3 weeks after sham or SNL surgery (n = 8 animals per group). Data are expressed as means ± SEM. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, compared with the respective baseline before drug injection (time 0; repeated measures ANOVA followed by the Dunnett’s post hoc test). D – F , lack of an effect of intrathecal injection of 100 μg JWH-133 on paw withdrawal thresholds tested using von Frey filaments ( D ), a pressure stimulus ( E ), and a heat stimulus ( F ) in rats 5 days after sham or SNL surgery (n = 7 rats per group). CB2, Type-2 cannabinoid receptors; SNL, spinal nerve ligation.

Article Snippet: The membrane was incubated with blocking solution (5% nonfat dry milk in Tris-buffered saline and 0.05% Tween 20 (TBST)) at 22 °C for 1 h and then incubated overnight at 4 °C with an anti-rabbit CB2 antibody (1:200 dilution, #101550, Cayman Chemicals) diluted with TBST containing 3% bovine serum albumin.

Techniques: Activation Assay, Injection, Ligation

Journal: The Journal of Biological Chemistry

Article Title: Cannabinoid CB2 receptors are upregulated via bivalent histone modifications and control primary afferent input to the spinal cord in neuropathic pain

doi: 10.1016/j.jbc.2022.101999

Figure Lengend Snippet: Nerve injury alters activating and repressive histone modifications at the Cnr2 promoter in the DRG. A , top , schematic representation of the rat Cnr2 gene exon and intron composition of different transcript variants. The coding sequence (starting with ATG) of the same 360 amino acid (aa) long CB2 protein (rCB2A) is localized solely within the common Exon 2 of both transcript variant 1 and variant 2. Transcript variant 3 (Exon 1–3), although 100% identical to Exon 2, encodes a longer (410 aa) CB2 protein (rCB2B). Bottom , alignment of rat CB2 protein isoforms with corresponding human (hCB2) and mouse (mCB2) isoforms shows the presence of a distinct C-terminal sequence in the rCB2B isoform, whereas the rCB2A isoform shares a high degree of similarity with hCB2 and mCB2. B , real-time qPCR analysis of Cnr2 mRNA levels using specific primers for each indicated Exon in rat DRG tissues 21 days after sham or SNL surgery. Data are expressed as mean ± SEM (n = 6 independent experiments; p < 0.0001, F(5,30) = 53.36). ∗∗∗ p < 0.001 compared with the sham group of Exon 1 (one-way ANOVA followed by Dunnett’s post hoc test). C , ChIP-qPCR quantification shows the enrichment of histone marks at -463 bp to -373 bp, -115 bp to -15 bp, -53 bp to +28 bp, and +117 bp to +219 bp regions flanking the transcription start site of the rat Cnr2 gene on chromosome 5. DRG tissues were obtained from rats 21 days after sham or SNL surgery and subjected to ChIP-qPCR to quantify the immunoprecipitated DNA using the antibodies against H3K9ac, H3K4me3, H3K9me2, H3K27me3, and pan-histone H3. The percentage of their enrichment at each indicated region on the Cnr2 promoter was normalized to the input sample and corrected with that of pan-histone H3 values. Rabbit IgG was used as a negative control in immunoprecipitations. The rat negative control primers spanning a gene desert on rat chromosome 3 was used to confirm the specificity of ChIP-qPCR. Data are expressed as means ± SEM (n = 6 independent experiments per group; DRG tissues from three rats were pooled as one sample). ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, compared with the sham group (two-tailed Student’s t test). CB2, Type-2 cannabinoid receptors; ChIP-qPCR, chromatin immunoprecipitation–quantitative PCR; DRG, dorsal root ganglion; SNL, spinal nerve ligation.

Article Snippet: The membrane was incubated with blocking solution (5% nonfat dry milk in Tris-buffered saline and 0.05% Tween 20 (TBST)) at 22 °C for 1 h and then incubated overnight at 4 °C with an anti-rabbit CB2 antibody (1:200 dilution, #101550, Cayman Chemicals) diluted with TBST containing 3% bovine serum albumin.

Techniques: Sequencing, Variant Assay, ChIP-qPCR, Immunoprecipitation, Negative Control, Two Tailed Test, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Ligation