711098

antibody from Invitrogen Antibodies

Targeting: CHRM1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

711098

Provider

Invitrogen Antibodies

Product name

CHRM1 Recombinant Polyclonal Antibody (14HCLC)

Antibody type

Polyclonal

Antigen

Synthetic peptide

Reactivity

Human

Host

Rabbit

Isotype

IgG

Antibody clone number

14HCLC

Vial size

100 µg

Concentration

0.5 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Normal and Pathological Tau Uptake Mediated by M1/M3 Muscarinic Receptors Promotes Opposite Neuronal Changes.

Morozova V, Cohen LS, Makki AE, Shur A, Pilar G, El Idrissi A, Alonso AD
Frontiers in cellular neuroscience 2019;13:403

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of Neuro-2a. The blots were probed with Anti-Muscarinic acetylcholine receptor M1 Recombinant Rabbit Polyclonal Antibody (Product # 711098, 1-2 µg/mL) 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). A 61 kDa band corresponding to Muscarinic acetylcholine receptor M1 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), 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® 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).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

For immunofluorescence analysis, IMR-32 cells were fixed and permeabilized for detection of endogenous Muscarinic acetylcholine receptor M1 using Anti- Muscarinic acetylcholine receptor M1 Recombinant Rabbit Polyclonal Antibody (Product # 711098, 2 µg/mL) and labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034, 1:2000). Panel a) shows representative cells that were stained for detection and localization of Muscarinic acetylcholine receptor M1 protein (green), Panel b) is stained for nuclei (blue) using SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). Panel c) represents cytoskeletal F-actin staining using Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d) is a composite image of Panels a, b and c clearly demonstrating membrane localization of Muscarinic acetylcholine receptor M1. Panel e) represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

FIGURE 1 Uptake of tau is cell-type specific. Recombinant tau and PH-Tau (400 ng/mL) were added to HEK293 and CHO cells (A) and to primary neuronal cultures (B) . Confocal analysis indicated that uptake occurred in HEK293 and neuronal cultures (white arrows) but not CHO cells and was blocked by pre-treatment with atropine. In the neuronal images, the boxed areas were zoomed 2.5x and converted (betaIII-tubulin, gray; tau, green). (C) Quantitation of tau uptake into neuronal cultures indicates that both tau (dark bars) and PH-Tau (light bars) are uptaken at similar levels. Pre-treatment with antagonists atropine and pirenzepine block the uptake of both proteins by about 80%. Conversely, AF-DX116 and PTX do not block uptake. (D) Left: Western blot analysis of cultured cells and neurons to analyze the presence of the M1 muscarinic receptor. Quantitation indicated that both HEK293 and neuronal cultures have a large amount of M1 muscarinic receptors compared to CHO cells. Right: Western blot analysis of HEK293 cells transfected with M1, M3, or M1/M3 muscarinic receptors to determine tau uptake. The introduction of the muscarinic receptors resulted in the significant uptake of tau into CHO cells. Scale bar: 20 mum. All experiments were performed three times with 10 images analyzed per experiment. ( * p < 0.05, ** p < 0.01, *** p < 0.001).