Western blotting was performed according to a previous report [28 (link)]. The Rabbit anti-VEGF (19003-1-AP), Rabbit anti-FLK1 (26415-1-AP), Rabbit anti-NFDUFS4 (15849-1-AP), Rabbit anti-alpha-SYN (10842-1-AP), Rabbit anti-TH (25859-1-AP), and Rabbit anti-GAPDH (10494-1-AP) primary antibodies were purchased from Proteintech (Wuhan, China). The Rabbit anti-P38MAPK (5140), Rabbit anti-ERK1/2 (4370), Rabbit anti-p-ERK1/2 (4695), Rabbit anti-SMAD3 (9523), Rabbit anti-p-SMAD3 (C25A9), Rabbit anti-caspase3 (9662) and Rabbit anti-ICAM1(4915) primary antibodies were purchased from Cell Signaling Technology (Danvers, USA). The FITC-labeled anti-Rabbit secondary antibodies were purchased from Cell Signaling Technology (Danvers, USA).
Rabbit anti gapdh 10494 1 ap
Manufactured by Proteintech
Sourced in China
Rabbit anti-GAPDH (10494-1-AP) is a primary antibody product used in various laboratory techniques, such as western blotting, immunohistochemistry, and immunofluorescence. The antibody specifically recognizes the GAPDH (Glyceraldehyde-3-Phosphate Dehydrogenase) protein, which is a commonly used housekeeping protein for normalization and control purposes in biological studies.
Automatically generated - may contain errors
Lab products found in correlation
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Rabbit anti caspase 3 9662, by Cell Signaling Technology (1 mentions)
Rabbit anti parp, by Cell Signaling Technology (1 mentions)
Rabbit anti pp4r1, by Merck Group (1 mentions)
Rabbit anti caspase 3, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase conjugated goat anti rabbit igg, by Santa Cruz Biotechnology (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
4 protocols using rabbit anti gapdh 10494 1 ap
1
Comprehensive Western Blot Protocol
2
CRISPR-mediated CBX3 Regulation in Colon Cancer
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Colon cancer cells were obtained from the tumor laboratory of the First Affiliated Hospital of Chongqing Medical University, China. The plasmid pX330-U6-Chimeric_BB-CBh-hSpCas9 was a gift from Feng Zhang (Addgene plasmid # 42230). Lentivirus plasmid pLKO.1 puro was a gift from Bob Weinberg (Addgene plasmid # 8453). Rabbit anti-CBX3 (A2248) was purchased from ABclonal Biotech Co (ABclonal, USA). The following antibodies were purchased from Abcam (Cambridge, MA): rabbit anti-CDK6 (ab124821), rabbit anti-p21 (ab109520). Rabbit anti-GAPDH (10494-1-AP) and rabbit anti-β-tubulin (10094-1-AP) were purchased from Proteintech Group, Inc. (Proteintech, China). Colon cancer tissue samples and matched adjacent normal tissues were derived from patients undergoing surgical procedures at the First Affiliated Hospital of Chongqing Medical University, China. All patients in the study provided written consent, and were approved by the Ethics Committee from the First Affiliated Hospital of Chongqing Medical University. Nude mice ages 4 to 5 weeks were purchased from Animal Experimental Center of Chongqing medical university. All animal care and handling procedures were handled in accordance with study protocols approved by the Ethics Committee of Chongqing Medical University.
3
Western Blot Analysis of Protein Targets
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Cells were scraped on ice, collected by centrifugation and lysed in 2×SDS sample buffer containing 4% SDS, 10% glycerol, 10 mM EDTA, and 100 mM Tris-HCl, pH 6.8 and collected by centrifugation. Subsequently, protein concentrations were determined by protein assay kit and total 30 µg proteins were separated on 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes at 300 mA for 2 hours. The blots were probed with primary antibodies overnight at 4℃ after being blocking in Tris buffered saline with Tween and 5% skim milk. Then the membrane was incubated with secondary antibody for 1 hour at room temperature. The primary antibodies were rabbit anti-PP4R1 (#HPA041089, 1:1,000; Sigma, St. Louis, USA), rabbit anti-caspase-3 (#9661, 1:500; Cell Signaling, Danvers, USA), rabbit anti-PARP (#9542, 1:1,000; Cell Signaling) and rabbit anti-GAPDH (#10494-1-AP, 1:500,000; Proteintech Group Inc., Chicago, USA) and horseradish peroxidase-conjugated goat anti-rabbit IgG (#SC-2054, 1:5,000; Santa Cruz Biotechnology, Santa Cruz, USA) was used as second antibody. The protein expression level was detected by enhanced chemiluminescence plus TM blotting system (Little Chalfont, Bucks, UK).
4
Western Blot Analysis of Protein Targets
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The WB analysis followed the protocols described in previous reports [22 (link)]. Briefly, tissue lysates were prepared with a RIPA buffer (Thermo Fisher Scientific, Waltham, MA, USA). The proteins (40 μg from each sample) were then separated on the 12% SDS-polyacrylamide gel, transferred to the PVDF membranes, and then blocked with 8% (w/v) skimmed milk in the Tris-buffered saline with a Tween 20 (TBST) buffer for 1 h. The membranes were washed thrice with the TBST buffer for 10 min each, followed by incubation with specific primary antibodies. The antibodies included: rabbit anti-GAPDH (10494-1-AP, Proteintech, Wuhan, China), rabbit anti-MTF1 (25383-1-AP, Proteintech), rabbit anti-SHP (ab96605, Abcam), and rabbit anti-DYNAMIN (18205-1-AP, Proteintech). We visualized the protein bands with enhanced chemiluminescent (ECL) and quantified them with Image-Pro Plus 6.0 software (Media Cybernetics, Rockville, MD, USA).
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