Antibodies against Necdin (ab18554), SGT1 (ab30931), BMAL1 (ab3350), HSP90 (ab13492), PER1 (ab136451), PER2 (ab180655), CRY1 (ab104736) were purchased from Abcam. Antibody against AVP (sc-390723) were obtained from Santa Cruz. Antibodies against Myc-tag (2276), Flag-tag (14793), HA-tag (3724), VIP (63269) were obtained from Cell Signaling Technology. Antibody against CRY2 were obtained from Invitrogen. Antibody against GRP (20073) were obtained from ImmunoStar. 17-AAG (HY-10211) and Geldanamycin (HY-15230) were purchased from MedChemExpress.
Ab104736
Manufactured by Abcam
Ab104736 is a primary antibody product offered by Abcam. It is a polyclonal antibody raised in rabbit and targets a specific protein. The antibody can be used in various immunoassay applications to detect the target protein.
Automatically generated - may contain errors
Lab products found in correlation
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Ab136451, by Abcam (1 mentions)
Ab180655, by Abcam (1 mentions)
Ab18554, by Abcam (1 mentions)
Ab13492, by Abcam (1 mentions)
Ab3350, by Abcam (1 mentions)
Ab30931, by Abcam (1 mentions)
Ecl detection kit, by Share-Bio (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
β actin, by Merck Group (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Na934, by Cytiva (1 mentions)
C57bl 6 magel2tm1stw j, by Jackson ImmunoResearch (1 mentions)
4 protocols using ab104736
1
Comprehensive Antibody Validation Protocol
2
Western Blot Analysis of Cell Signaling Proteins
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Exponential phase cells were washed by PBS twice and added with RIPA buffer, then collected using a cell scraper. Cell lysates were vibrated 10s every 5min for 30 min at 0°C and centrifuged for 10 min (12,000 rpm, 4°C), and then the supernatants were transferred into clean EP tubes. Protein concentration was quantified using a BCA Protein Assay Kit according to the manufacturer's instructions (Beyotime, China). The supernatants (10μg protein) were subjected to SDS-PAGE gel, after which the proteins were transferred to PVDF membranes (Millipore, USA), which were then blocked with 5% non-fat dried milk for 2 hours at room temperature. The membranes were then probed with primary antibodies for Cry1 (ab104736, Abcam), Akt1 (sc-1618, Santa cruz), p-Akt (4060, CST), MDM2 (sc-965, Santa cruz), p-MDM2 (35215, CST), P53 (sc-126, Santa cruz), P21 (2947, CST), Cdk2 (sc-163, Santa cruz), Cyclin E (sc-247, Santa cruz), Cyclin A (sc-751, Santa cruz) and β-actin (a1978, Sigma) at 4°C overnight. The membranes were washed three times in TBST, and then incubated with goat anti-rabbit or goat anti-mouse IgG-horseradish peroxidase secondary antibody (7074, 4410, CST, USA) at 37°C for 2 h. The ECL Detection kit (Share-bio, China) was used for detection and photography. Images were read with a Fluor Chem E system (Proteinsinple, USA). The assays were performed in triplicate.
3
Cryptochrome1 Expression in Magel2-Null Mice
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All animal studies were conducted in accordance with the Canadian Council on Animal Care Guidelines and Policies with approval from the Animal Care and Use Committee: Health Sciences for the University of Alberta. Adult Magel2+m/-p (Magel2-null) and control littermate Magel2+m/+p mice (Jackson Laboratories C57BL/6-Magel2tm1Stw/J, stock 009062) were genotyped by PCR of spare tissue. Postnatal day 10 mouse pups were euthanized and brains removed then the cortex and hypothalamus were dissected and snap frozen in liquid nitrogen. The tissues were ground using pre-chilled disposable plastic pestles and resuspended in 500 μl of 2X modified sample buffer with Complete Mini Protease Inhibitor. The tissues were sonicated on ice (3x for 5 s each with 5 s pauses), incubated at 65°C for 5 min, and centrifuged 10 min at 20000 g at room temperature, and supernatants retained. For immunoblot analysis, 20 μg of protein was subjected to SDS-PAGE and immunoblotting as described above, then blots were probed with rabbit polyclonal anti-Cryptochrome1 (Abcam ab104736, 1:2000) followed by HRP-linked donkey anti-rabbit IgG (Amersham Pharmacia Biotech #NA934, 1:5000).
4
Quantifying Liver Protein Expression
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Liver cytoplasmic extracts were prepared as described previously (Jouffe et al. 2013 (link)). Protein extract concentrations were quantified using a BCA protein assay kit (Thermo Fisher Scientific), and 20 µg of liver protein extract was resolved by SDS-PAGE using standard procedures. Densitometry analyses of the blots were performed using the ImageJ software. Naphtol blue and black staining of the membranes was used as a loading control and served as a reference for normalization of the quantified values. CRY1 antibody (1/500) was from Abcam (ab104736).
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