Frozen specimens from rodents were homogenized in 100 μl lysis buffer containing a mixture of proteinase and phosphatase inhibitors and then centrifuged at 15 000 rpm for 15 min at 4°C. The protein concentration was determined using a BCA Protein Assay Kit (Pierce, Rockford, IL). A total of 50 μg of protein was resolved on 12% precasted SDS‐PAGE gels, then transferred to a polyvinylidene fluoride (PVDF) membrane (Millipore, Bedford, MA). The PVDF membrane was blocked with 5% non‐fat milk in PBS containing 0.1% Tween 20 for 2 h at room temperature and then incubated overnight at 4°C with primary antibodies. The following antibodies were used in this study: GAPDH, anti TNF‐α, anti‐ASIC1, and anti‐ASIC3 antibody (1:1000, abcam, USA). After washing with Tris‐Buffered Saline, 0.1% Tween 20 Detergent (TBST), the blots were incubated for 2 h at room temperature with HRP‐conjugated secondary antibody (1:5000; Amersham Biosciences, San Francisco, CA, USA), visualized by using Electro‐Chemi‐Luminescence (ECL) chemiluminescent detection system (Amersham Biosciences).
Anti asic1
Manufactured by Abcam
Sourced in United States
Anti‐ASIC1 is a primary antibody that recognizes the Acid‐Sensing Ion Channel 1 (ASIC1) protein. ASIC1 is a subunit of the acid‐sensing ion channel, which plays a role in the detection of acidic pH changes in the body. This antibody can be used in various research applications to study the expression and localization of ASIC1 in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Anti asic3 antibody, by Abcam (2 mentions)
Hrp conjugated secondary antibody, by Cytiva (2 mentions)
Electro chemi luminescence chemiluminescent detection system, by Cytiva (2 mentions)
Anti tnf α, by Abcam (2 mentions)
Pvdf membrane, by Merck Group (2 mentions)
Gapdh, by Abcam (2 mentions)
Anti asic3, by Abcam (2 mentions)
Alexa 488 conjugated goal anti rabbit igg, by Jackson ImmunoResearch (2 mentions)
4 protocols using anti asic1
1
Western Blot Analysis of Protein Expression
2
Western Blot Analysis of Protein Expression
3
Dual ASIC1 and ASIC3 Immunofluorescence
4
Immunofluorescence Analysis of ASIC Channels
Check if the same lab product or an alternative is used in the 5 most similar protocols
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After the rats were sacrificed, the chest was opened, and the ascending aorta was then infused with ice‐cold saline and 4% paraformaldehyde. The T3–T5 DRGs were removed and incubated with 4% paraformaldehyde for 3 h at room temperature and then replaced with 30% sucrose for 24 h at 4°C. The DRGs were then embedded in Histoprep and were cut at a thickness of 10 μm on a cryostat.
For double immunofluorescence, the DRG sections were incubated with a mixture of anti‐ASIC1 (1:1000, abcam, USA), monoclonal neuronal‐specific nuclear protein (NeuN) (1:500, Millipore, USA), and anti‐ASIC3 (1:1000, abcam, USA) overnight at 4°C. The sections were washed with PBS and then incubated with Alexa 488‐conjugated goal anti‐rabbit IgG (1:200; Jackson ImmunoResearch Laboratories, USA) for 2 h at room temperature.
For double immunofluorescence, the DRG sections were incubated with a mixture of anti‐ASIC1 (1:1000, abcam, USA), monoclonal neuronal‐specific nuclear protein (NeuN) (1:500, Millipore, USA), and anti‐ASIC3 (1:1000, abcam, USA) overnight at 4°C. The sections were washed with PBS and then incubated with Alexa 488‐conjugated goal anti‐rabbit IgG (1:200; Jackson ImmunoResearch Laboratories, USA) for 2 h at room temperature.
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