QM5 cells were maintained in medium 199 with Earle’s salts containing 100 U/mL of penicillin and streptomycin and 10% heat-inactivated fetal bovine serum. Monoclonal mouse anti-FLAG (Sigma-Aldrich) and rabbit anti-Myc (Sigma-Aldrich) antibodies, horseradish peroxidase (HRP)-conjugated goat anti-rabbit (Santa Cruz) and goat-anti mouse (Santa Cruz) antibodies, Alexa Fluor 488-conjugated goat anti-mouse (Invitrogen) and Alexa Fluor 555-conjugated goat anti-rabbit (Invitrogen), Sulfo-NHS-LC-Biotin (Thermo Scientific), maliemide-PEG2-biotin (Thermo Scientific), neutravidin agarose resin (Thermo Scientific), and polyethylimine (PEI, Polysciences Inc.) were purchased from the indicated commercial sources.
Horseradish peroxidase hrp conjugated goat anti rabbit
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Horseradish peroxidase (HRP)-conjugated goat anti-rabbit is a secondary antibody that binds to rabbit primary antibodies. HRP is an enzyme that can catalyze colorimetric or chemiluminescent reactions, allowing for the detection of the bound primary antibody.
Automatically generated - may contain errors
Lab products found in correlation
Mouse monoclonal anti flag, by Merck Group (2 mentions)
Alexa fluor 488 conjugated goat anti mouse, by Thermo Fisher Scientific (2 mentions)
Neutravidin agarose resin, by Thermo Fisher Scientific (2 mentions)
Sulfo nhs lc biotin, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 555 conjugated goat anti rabbit, by Thermo Fisher Scientific (1 mentions)
Rabbit anti myc, by Merck Group (1 mentions)
Alexa fluor 647 conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions)
Rabbit anti c myc, by Merck Group (1 mentions)
Maleimide peg2 biotin, by Thermo Fisher Scientific (1 mentions)
Methyl β cyclodextrin, by Merck Group (1 mentions)
Enhanced chemiluminescence, by Merck Group (1 mentions)
Polyvinylidene fluoride membrane, by Merck Group (1 mentions)
Anti vegf rabbit pab, by Santa Cruz Biotechnology (1 mentions)
Anti gapdh mouse mab, by Santa Cruz Biotechnology (1 mentions)
Anti phospho akt, by Cell Signaling Technology (1 mentions)
Anti akt rabbit pab, by Cell Signaling Technology (1 mentions)
Crystal violet solution, by Merck Group (1 mentions)
Anti p p70s6k1, by Cell Signaling Technology (1 mentions)
Anti p70s6k1, by Cell Signaling Technology (1 mentions)
Anti p ampk, by Cell Signaling Technology (1 mentions)
5 protocols using horseradish peroxidase hrp conjugated goat anti rabbit
1
Cell Culture and Antibody Labeling
2
ARV p10 Characterization Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
QM5 and Vero cells were grown and maintained as previously described (Corcoran and Duncan, 2004). Rabbit antisera generated against full-length ARV p10 or ARV p10 endodomain were previously described [43] (link), [78] (link). Monoclonal mouse anti-FLAG (Sigma-Aldrich) and rabbit anti-c-myc (Sigma-Aldrich) antibodies, horseradish peroxidase (HRP)-conjugated goat anti-rabbit (Santa Cruz) and goat-anti mouse (Santa Cruz) antibodies, Alexa Fluor 488-conjugated goat anti-mouse (Invitrogen) and goat anti-rabbit (Invitrogen), antibodies, Alexa Fluor 647-conjugated goat anti-rabbit IgG (Invitrogen), maleimide-PEG2-biotin (Thermo Scientific), neutravidin agarose resin (Thermo Scientific), and methyl-β-cyclodextrin (Sigma-Aldrich) were purchased from the indicated commercial sources.
3
Western Blot Analysis of Key Signaling Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Proteins were separated by 8% and 10% SDS polyacrylamide gels and electrophoretically transferred onto polyvinylidene fluoride membranes (Millipore, Billerica, MA, USA). The membranes were blocked overnight with 5% non-fat dried milk and incubated overnight with antibodies. Anti-Kidins220 rabbit polyclonal antibody (PAb) (Cat No. ab97345) was purchased from Abcam Inc (Abcam, Cambridge, UK). Anti-PI3K rabbit monoclonal antibody (MAb) (Cat No. ♯4257), anti-phospho-PI3K rabbit PAb (Cat No. ♯4228), anti-AKT rabbit PAb (Cat No. ♯9272) and anti-phospho-AKT (Cat No. ♯4060) were all purchased from Cell Signaling Technologies (Beverly, MA, USA). Anti-VEGF rabbit PAb (Cat No. sc-152), anti-GAPDH mouse MAb (Cat No. sc-47724), anti-a-Tubulin mouse MAb (Cat No. sc-23948) and horseradish peroxidase (HRP) conjugated goat anti-rabbit (Cat No. sc-2004) or anti-mouse (Cat No. sc-2005) secondary antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The signals were detected by enhanced chemiluminescence (Millipore).
4
Cellular Responses to Pharmacological Agents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fetal bovine serum (FBS), cell culture media, penicillin/streptomycin, and all other reagents used for cell culture studies were purchased from Welgene (Gyeongsan, Korea). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT), 4,6-diamidino-2-phenylindole (DAPI), 5-Fu, rhodamine 123 (Rh123), CoCl2, compound C, crystal violet solution, and other reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA). Anti-Akt, anti-p-Akt, anti-p-AMPK, anti-Bcl-2, anti-caspase-3, anti-cleaved caspase-3, anti-p-ERK, anti-PARP, anti-mTOR, anti-p-mTOR, anti-p70S6K1, anti-p-p70S6K1, and anti-p-4E-BP1 antibodies were supplied by Cell Signaling Technologies (Danvers, MA, USA). Anti-β-actin and anti-vascular endothelial growth factor (VEGF) antibodies, horseradish peroxidase (HRP)-conjugated goat anti-rabbit, and goat anti-mouse IgGs were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-hypoxia-inducible factor-1α (HIF-1α) antibody was obtained from BD Biosciences (SanJose, CA, USA). Dimethyl sulfoxide (DMSO) was purchased from AppliChem (Darmstadt, Germany) and Junsei Chemical Co. (Tokyo, Japan).
5
Biomarker Evaluation in Cell Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For biomarker evaluation, primary antibodies used against HGF (clone 7-2, Novus Biologicals, NBP1-19182), p-Met (Tyr 1234/1235) (clone D26, Cell Signaling Technology, Cat# 3077s), Gab1 (clone H-198, Santa Cruz Biotechnology, sc-9049), pAkt (Ser-473, Santa Cruz Biotechnology, sc-7985-R), NF-κB p50 (NF-κB1 or subunit p50, NLS, Santa Cruz Biotechnology, sc-114), COX-2 (Santa Cruz Biotechnology, sc-7951), SDF-1α (clone FL-93, Santa Cruz Biotechnology, sc-28876), CXCR4 (clone H-118, Santa Cruz Biotechnology, sc-9046), β-actin (Santa Cruz Biotechnology, sc-7210) and horseradish peroxidase (HRP)-conjugated goat anti-rabbit (Santa Cruz Biotechnology, sc-2004) secondary antibody were used in this study. For detection of chromogenic signal, the immunohistochemistry assay kit (DAB150: IHC Select HRP/DAB Tests, Merck Millipore, Germany) was used. For Western blot analysis, chemiluminescent detection was performed by using Clarity Western ECL Substrate (Bio-Rad).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!