The sources of antibodies and the dilutions used in experiments were as follows: rabbit anti-β-PIX (Millipore 07-1450, Darmstadt, Germany; dilution for Western blotting: 1/1000; dilution for immunofluorescence: 1/500); mouse anti-paxillin (BD Bioscience 610052, San Jose, California, USA; dilution for immunofluorescence: 1/1000); mouse anti-GAPDH (GeneTex GTX627408, Hsinchu, Taiwan; dilution for Western blotting: 1/1000); DNase I (Sigma-Aldrich SAB2702031, Saint Louis, Missouri, USA; dilution for immunofluorescence: 1:1000); Alexa Fuor 488-antimouse IgG (Jackson ImmunoResearch 715-546-151/J1, West Grove, Philadelphia, USA; dilution for immunofluorescence: 1/300); Alexa Fuor 568-antimouse IgG (Invitrogen A11031; dilution for immunofluorescence: 1/300); Alexa Fuor 568-anti-rabbit IgG (Invitrogen A11036; dilution for immunofluorescence: 1/300); Alexa Fuor 647-antimouse IgG (Jackson ImmunoResearch 715-606-151/J1; dilution for immunofluorescence: 1/300). Alexa Fluor 488 phalloidin (A12379; dilution for immunofluorescence: 1/400) and Alexa Fluor 568 phalloidin (A12380; dilution for immunofluorescence: 1/400) were obtained from Invitrogen. HRP-AffiniPure goat antimouse IgG (115-035-174; dilution for Western blotting: 1/5000) and HRP-AffiniPure goat anti-rabbit IgG (211-032-171; dilution for Western blotting: 1/5000) were obtained from Jackson ImmunoResearch.
Mouse anti gapdh
Manufactured by GeneTex
Sourced in United States
Mouse anti-GAPDH is a primary antibody that recognizes the GAPDH (Glyceraldehyde-3-Phosphate Dehydrogenase) protein. GAPDH is a key enzyme involved in glycolysis and is commonly used as a loading control or reference protein in various experimental techniques.
Automatically generated - may contain errors
Lab products found in correlation
Chemidoc xrs imaging acquiring system, by Bio-Rad (5 mentions)
Clarity western ecl substrate chemiluminescence kit, by Bio-Rad (5 mentions)
Image lab software v 6, by Bio-Rad (4 mentions)
Rabbit anti erk1 2, by Cell Signaling Technology (3 mentions)
Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (2 mentions)
Mouse anti paxillin, by BD (2 mentions)
Hrp affinipure goat anti mouse igg, by Jackson ImmunoResearch (2 mentions)
Cell extraction buffer, by Thermo Fisher Scientific (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Rabbit anti p erk1 2 thr202 tyr204, by Cell Signaling Technology (2 mentions)
Dnase 1, by Merck Group (1 mentions)
Alexa fluor 568 phalloidin, by Thermo Fisher Scientific (1 mentions)
Mouse anti β actin, by Santa Cruz Biotechnology (1 mentions)
Mouse anti nr2b, by Abcam (1 mentions)
Mouse anti β actin, by Merck Group (1 mentions)
Mouse anti glua2, by Merck Group (1 mentions)
Mouse anti psd95, by Thermo Fisher Scientific (1 mentions)
Rabbit anti foxg1, by Abcam (1 mentions)
Dc protein assay, by Bio-Rad (1 mentions)
Irdye 800cw goat anti rabbit igg, by LI COR (1 mentions)
17 protocols using mouse anti gapdh
1
Antibodies and Dilutions for Immunofluorescence and Western Blotting
2
Protein Quantification and Western Blotting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell protein lysate was obtained using cell extraction buffer (Life Technologies, Carlsbad, CA, USA) followed by incubation for 30 min, at 4 °C, on a shaker. After centrifugation, the supernatant was collected and submitted to protein quantification by a BCA Protein Assay Kit. SDS-Page and Western blots were performed as described using stain-free gels [15 (link)]. Membranes were incubated with mouse anti-GAPDH, mouse anti-ILK [N1C1] (GeneTex, Irvine, USA), mouse anti-β-actin, mouse anti-β1-integrin P5D2, rabbit anti-CTR1 [FL190], goat anti-MRP2 [H17] (Santa Cruz Biotechnology), as well as goat anti-rabbit, donkey anti-goat and anti-mouse IgG kappa binding protein IgG HRP-conjugated (Santa Cruz Biotechnology) diluted in 1% BSA solution. Western blots were quantified via chemiluminescence using a Clarity Western ECL substrate chemiluminescence kit (BioRad Laboratories GmbH, Munich, Germany). Besides the loading control GAPDH, we also used stainfree total protein normalization. Membranes were photographed and analyzed using a ChemiDoc XRS+ imaging acquiring system (BioRad) and Image Lab software v. 6.0 (BioRad).
3
Antibody Characterization for Western Blotting
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used for western blotting: mouse anti-Synaptotagmin (1:250; Abcam, Cambridge, UK); rabbit anti-GluA1 (1:1000, Thermo Fisher Scientific, Dreieich, Germany); mouse anti-GluA2 (1:1000; Millipore, Schwalbach, Germany); mouse anti-NR1 (1:1000; BD Biosciences, San Jose, CA); rabbit anti-NR2A (1:1000; Abcam, Cambridge, UK); mouse anti-NR2B (1:1000; Abcam, Cambridge, UK); mouse anti-PSD-95 (1:1000; Thermo Fisher Scientific, Dreieich, Germany); mouse anti-β-actin (1:5000; Sigma, Taufkirchen, Germany); mouse anti-GAPDH (1:5000; GeneTex, Irvine, CA), peroxidase-conjugated goat anti-mouse and goat anti-rabbit (1:15,000; Dianova, Hamburg, Germany).
4
Foxg1 Expression in Neocortex Development
5
Comprehensive Antibody Characterization of Tubulin Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following primary and secondary antibodies were used: mouse anti-Tubb1 (Novus; #NBP2-46245; IF 1:200); rabbit anti-Tubb2 (Abcam; #ab179512; IF 1:200); mouse anti-Tubb3 (Biolegend; #801202; IF 1:1,000; WB 1:5,000); mouse anti-Tubb4 (Novus; #NB-120-11315; IF 1:100); mouse anti-Tubb5 (Novus; #NB-120-11312; IF 1:200); mouse anti-panTuba (Sigma; #T9026; IF 1:1,000); rabbit anti-panTubb (Abcam; #ab151318; IF 1:200); rat anti-panTuba (Origene; #SM568P; IF 1:500); rabbit anti-Polyglutamate chain (PolyE) (AdipoGen; #AG-25B-0030; IF 1:1000); mouse anti-Glutamate Receptor 2 (Millipore; #MAB397; WB 1:1000); mouse anti-Homer1 (Synaptic Systems; #160011; WB 1:1000); mouse anti-GAPDH (GeneTex; #GTX28245; WB 1:5000); mouse anti-gamma-adaptin (BD Bioscience; #610386; WB 1:5,000); donkey anti-mouse HRP-conjugated (Dianova; #715-036-151; WB 1:10,000); Cy3 donkey anti-mouse IgG (Dianova; #715–165-150; IF 1:500); Cy3 donkey anti-rabbit (Dianova; #711-166-152; IF 1:500). DNA stainings were performed with: DAPI (Thermo Fischer; #D1306: 1:1,000). IRDye 680RD goat anti-mouse IgG (LI-COR, NE, USA, 926-68070, WB 1:10,000); IRDye 800CW goat anti-rabbit IgG (LI-COR, NE, USA, 926-32211, WB 1:10,000).
6
Western Blot Analysis of E-cadherin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were washed twice to exclude dead cells. Then, cells were lysed using extraction buffer (Life Technologies, Carlsbad, CA, USA), supplemented with phenylmethanesulfonylfluoride (0.1 mM PMSF) (Life Technologies, Carlsbad, CA, USA) and a protease inhibitor cocktail (1 μg/mL aprotinin, 1 μg/mL leupeptin) (Life Technologies, Carlsbad, CA, USA), according to manufacturer’s instructions. Cells were then centrifuged and supernatant was collected and submitted to protein quantification by Pierce™ BCA Protein Assay Kit (Thermo Fisher Scientific Inc, Waltham, MA USA). SDS-Page and Western Blot were performed, as described in [54 (link)]. Membranes were incubated with rabbit anti- E-cadherin (Santa Cruz Biotechnology, Dallas, TX, USA), mouse anti-GAPDH (GeneTex, Irvine, CA, USA), and goat anti-rabbit and donkey anti-mouse IgG HRP-conjugated (Santa Cruz Biotechnology, Dallas, TX, USA) diluted in 1% BSA solution. Western Blot was quantified via chemiluminescence using Clarity Western ECL substrate chemiluminescence kit (BioRad). Membranes were photographed and quantified using ChemiDoc XRS+ imaging acquiring system (BioRad) and Image Lab software (BioRad).
7
Immunofluorescence and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse anti-SAS6, rabbit anti-cortactin, and mouse anti-γ-tubulin: Santa Cruz; rabbit anti-STIL and mouse anti-TRIO: Abcam; rabbit anti-GEF-H1 and rabbit anti-integrin β1 (total form), mouse anti-GAPDH, rabbit anti-GOLPH2, rabbit anti-β-actin, rabbit anti-paxillin, mouse anti-γ-tubulin, and rabbit anti-NMIIA: GeneTex; mouse anti-α-tubulin and mouse anti-acetylated tubulin: Sigma-Aldrich; rat anti-α-tubulin: ABD; mouse anti-paxillin: BD; mouse anti-Rac1 and mouse anti-integrin β1 (active form): Millipore; rabbit anti-TRIO, Alexa Fluor 488 phalloidin, Alexa Fluor 488-anti-rat IgG, Alexa Fluor 488-anti-rabbit IgG, Alexa Fluor 488-anti-mouse IgG, Alexa Fluor 568-anti-mouse IgG, Alexa Fluor 568-anti-rabbit IgG, Alexa Fluor 680-anti-rabbit IgG, and DAPI: Thermo Fisher; and HRP-AffiniPure mouse anti-rabbit IgG and HRP-AffiniPure goat anti-mouse IgG: Jackson ImmunoResearch.
8
Immunostaining and Western Blot Protocols
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For immunostaining, the primary antibodies used were mouse anti-CD63 (Developmental Studies Hybridoma Bank, Cat. No. H5C6), mouse anti-clathrin LC (Biolegend, Cat. No. MMS-423P), mouse anti-caveolin (Abcam, Cat. No. ab17052), LAMP1 (Developmental Studies Hybridoma Bank, Cat. No. H4A3), mouse anti-EEA1 (BD Biosciences, Cat. No. 610456), and rabbit anti-NanoLuc (Promega). All primary antibodies for immunostaining experiments were used at 1 μg/mL. Secondary antibodies used were donkey anti-mouse Alexa Fluor 546 (Life Technologies) and donkey anti-rabbit Alexa Fluor 488 (Life Technologies). All secondary antibodies for immunostaining experiments were used at 8 μg/mL.
For western blot, primary antibodies used were rabbit anti-NanoLuc (Promega), and mouse anti-GAPDH (GeneTex, Cat. No. GT239), diluted to 0.74 μg/mL and 1 μg/mL, respectively, in Odyssey blocking buffer (LI-COR) with 0.1% Tween-20. The secondary antibodies were a donkey anti-mouse IRDye 680RD (LI-COR) and a donkey anti-rabbit IRDye 800 (LI-COR) and were diluted 1:3,000 in Odyssey blocking buffer with 0.1% Tween-20.
For western blot, primary antibodies used were rabbit anti-NanoLuc (Promega), and mouse anti-GAPDH (GeneTex, Cat. No. GT239), diluted to 0.74 μg/mL and 1 μg/mL, respectively, in Odyssey blocking buffer (LI-COR) with 0.1% Tween-20. The secondary antibodies were a donkey anti-mouse IRDye 680RD (LI-COR) and a donkey anti-rabbit IRDye 800 (LI-COR) and were diluted 1:3,000 in Odyssey blocking buffer with 0.1% Tween-20.
9
Fibulin-3 Regulation via KISS1R-MMP9 Pathway
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Experiments were performed previously as described [37 (link), 39 (link)]. Cells were lysed using RIPA buffer and protein was separated by SDS-PAGE. Protein (50 µg) levels were quantified using antibodies raised against human proteins: rabbit anti-fibulin-3 (1:1000; Abcam), rabbit anti-KISS1R (1:4000, Abcam), rabbit anti-MMP-9 (1:500; Abcam), rabbit anti-ERK1/2 (1:1000; Cell Signaling), rabbit anti-phoshpo-ERK1/2 (1:2000; Cell Signaling), rabbit anti-AKT (1:1000; Cell Signaling), rabbit anti-phospho-AKT (1:1000; Cell Signaling). β-Actin or glyceraldehyde 3-phosphate dehydrogenase (GAPDH) expression was used as a loading control and was examined using rabbit anti-Actin (1:5000; GeneTex Inc.) or mouse anti-GAPDH (1:3000; GeneTex Inc.). After 1-hour incubation with horseradish peroxidase (HRP)-conjugated secondary antibodies, rabbit (1:2500, GE Healthcare) or mouse (1:2500, GE Healthcare), the proteins were visualized using SuperSignal West Dura Extended Duration Substrate (Thermo Scientific) and a VersaDoc Imaging System (Bio-Rad).
To determine the effect of KP-10 treatment on fibulin-3 expression, cells were treated with KP-10 (100 nM) (Calbiochem) in FBS-supplemented media for 72 hours, where applicable.
To determine the effect of KP-10 treatment on fibulin-3 expression, cells were treated with KP-10 (100 nM) (Calbiochem) in FBS-supplemented media for 72 hours, where applicable.
10
Immunoblotting for Sun Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Antibodies used for immunoblotting were as follows: mouse antitubulin (1:5,000; Sigma-Aldrich), mouse anti-GAPDH (1:5,000; Genetex), rabbit anti-Sun1 (1:1,000; Novus Biologicals), and rabbit anti-Sun2 (1:1,000; Genetex). Alexa Fluor 680– or 800–conjugated secondary antibodies (Thermo Fisher Scientific) were used at 1:10,000. Primary and secondary antibodies were incubated on Western blots for 1 h each. Western blot bands were quantified using a fluorescent imager (Odyssey; LI-COR Biosciences) and ImageJ. The integrated density of the bands was background subtracted and normalized to tubulin or GAPDH before comparison.
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!