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Western blotAntibody data
- Antibody Data
- Antigen structure
- References [1]
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- Validations
- Western blot [1]
- Immunocytochemistry [1]
- Other assay [1]
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- Product number
- 711098 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- CHRM1 Recombinant Polyclonal Antibody (14HCLC)
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with Monkey, Pig, House and Rat
- Antibody clone number
- 14HCLC
- Concentration
- 0.5 mg/mL
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
Frontiers in cellular neuroscience 2019;13:403
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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).
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.
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).
