The total protein of primary osteoblasts was extracted using RIPA lysis buffer (Beyotime, China) and quantified by BCA assay (Beyotime, China). Equal amounts of proteins (100 μg) were separated via BeyoGel™ Plus PAGE (Beyotime, China) and then transferred to a PVDF membrane (Millipore, USA). After transferring, the membranes were blocked with 5% fat-free milk for 1 h. The membranes were incubated with primary antibodies (Bax, ab32503; Caspase-3, ab32351; and cleaved-Caspase-3, ab32042) purchased from Abcam (USA) and GAPDH, 10494-I-AP purchased from Proteintech (China) at 4°C overnight. The membranes were incubated with second antibodies (goat anti rabbit IgG HRP SE134, goat anti mouse IgG HRP SE131, Solarbio, China) at 37°C for 1 h. The ECL system (CLINX, China) was used for exposing protein bands. The intensity of the bands was analyzed using Image Lab (version 3.0, Bio-Rad, USA).
Goat anti rabbit igg hrp se134
Manufactured by Solarbio
Sourced in China
Goat anti rabbit IgG HRP SE134 is a laboratory reagent used in immunoassays and other applications that require the detection of rabbit immunoglobulin G (IgG). It consists of goat-derived antibodies that are specific to rabbit IgG and are conjugated with horseradish peroxidase (HRP), an enzyme that can be used for colorimetric or chemiluminescent detection.
Automatically generated - may contain errors
Lab products found in correlation
Pvdf membrane, by Merck Group (4 mentions)
Goat anti mouse igg hrp se131, by Solarbio (4 mentions)
Ripa lysis buffer, by Beyotime (3 mentions)
Bca assay, by Beyotime (3 mentions)
Image lab version 3, by Bio-Rad (3 mentions)
Ecl system, by Clinx (3 mentions)
Ab32042, by Abcam (3 mentions)
Ab32351, by Abcam (3 mentions)
Beyogel plus page, by Beyotime (3 mentions)
Cleaved caspase 3, by Abcam (2 mentions)
Caspase 3, by Abcam (2 mentions)
Bcl2 associated x (bax), by Abcam (2 mentions)
Ab32503, by Abcam (2 mentions)
Ecl reagent, by Solarbio (1 mentions)
A20798, by ABclonal (1 mentions)
A18196, by ABclonal (1 mentions)
Ripa buffer, by Solarbio (1 mentions)
P erk, by ABclonal (1 mentions)
Sab2900314, by Merck Group (1 mentions)
A19607, by ABclonal (1 mentions)
4 protocols using goat anti rabbit igg hrp se134
1
Western Blot Analysis of Osteoblast Proteins
2
Protein Quantification and Western Blot Analysis
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Total protein was extracted using RIPA buffer (Solarbio). Quantification was performed with the BCA protein concentration assay kit (Solarbio). Approximately 20 µg total protein was passed through 10% SDS polyacrylamide gels, and then transferred to PVDF membranes (Millipore, USA). The membranes were incubated with the primary antibodies (CBLC, 1:1000, SAB2900314, Sigma; ERK, 1:500, 13–6200, Invitrogen; p-ERK, 1:1000, A18196, Abclonal; ABI1, 1:10,000, 66,609–1-Ig, Proteintech; CDK1, 1:1000, A22347, ABclonal; cyclin B, 1:2000, 55,004–1-AP, Proteintech; E-cadherin, 1:1000, A20798, ABclonal and Vimentin, 1:1000, A19607, ABclonal) at 4 °C overnight, followed by incubation with the corresponding secondary antibodies (1:3000, goat anti-rabbit IgG-HRP, SE134, Solarbio; 1:3000, or goat anti-mouse IgG-HRP, SE131, Solarbio) in the dark at room temperature for 60 min. The blots were developed with enhanced chemiluminescence (ECL) reagent (Solarbio), and data analysis was performed by Gel-Pro-Analyzer software.
3
Protein Analysis of Primary Osteoblasts
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The total protein of primary osteoblasts was extracted using RIPA lysis buffer (Beyotime, China) and quanti ed by BCA assay (Beyotime, China). Equal amounts of proteins (100μg) were separated via BeyoGel™ Plus PAGE (Beyotime, China) and then transferred to a PVDF membrane (Millipore, USA). After transferring, the membranes were blocked with 5% fat-free milk for 1h. The membranes were incubated with primary antibodies (Bax, ab32503, Caspase-3, ab32351, cleaved-Caspase-3, ab32042), which were purchased from Abcam (USA) and GAPDH, 10494-I-AP which was obtained from Proteintech (China) at 4°C overnight. The membranes were incubated with the second antibodies (Goat anti rabbit IgG HRP SE134, Goat anti mouse IgG HRP SE131, Solarbio, China) at 37°C for 1h. The ECL system (CLINX, China)
was used for exposing protein bands. The intensity of the bands was analyzed using Image lab (version 3.0, Bio-Rad, USA).
was used for exposing protein bands. The intensity of the bands was analyzed using Image lab (version 3.0, Bio-Rad, USA).
4
Protein Analysis of Primary Osteoblasts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The total protein of primary osteoblasts was extracted using RIPA lysis buffer (Beyotime, China) and quanti ed by BCA assay (Beyotime, China). Equal amounts of proteins (100μg) were separated via BeyoGel™ Plus PAGE (Beyotime, China) and then transferred to a PVDF membrane (Millipore, USA). After transferring, the membranes were blocked with 5% fat-free milk for 1h. The membranes were incubated with primary antibodies (Bax, ab32503, Caspase-3, ab32351, cleaved-Caspase-3, ab32042), which were purchased from Abcam (USA) and GAPDH, 10494-I-AP which was obtained from Proteintech (China) at 4°C overnight. The membranes were incubated with the second antibodies (Goat anti rabbit IgG HRP SE134, Goat anti mouse IgG HRP SE131, Solarbio, China) at 37°C for 1h. The ECL system (CLINX, China)
was used for exposing protein bands. The intensity of the bands was analyzed using Image lab (version 3.0, Bio-Rad, USA).
was used for exposing protein bands. The intensity of the bands was analyzed using Image lab (version 3.0, Bio-Rad, USA).
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