Total cell extracts were resolved on a SDS–polyacrylamide gel and blotted on Amersham Hybond P0.45 PVDF membrane (GE Healthcare Life Science). Membranes were blocked by non‐fat dry milk Blotting‐Grade Blocker (Bio‐Rad) 5% in PBS 0.1% Tween‐20 (Sigma‐Aldrich), incubated with primary antibodies overnight at 4°C, washed and hybridized for 1 h at RT using the appropriate secondary antibody. The following antibodies were used: anti‐p27 (BD Biosciences, 610241); anti‐p63 (Abcam, ab735); anti‐p63α (Cell Signaling, CS13109); anti‐KLF4 (R&D System, 12173S); anti‐K10 (BioLegend, PRB‐159P); anti‐LOR (BioLegend, PRB‐145P); anti‐β‐actin AC‐15 (Sigma, A5441); goat anti‐mouse HRP Conjugate (Bio‐Rad, 170‐5047); goat anti‐rabbit HRP Conjugate (Bio‐Rad, 170‐6517); and bovine anti‐goat HRP conjugate (Santa Cruz Biotechnology, SC‐2350).
Goat anti rabbit hrp conjugate
Manufactured by Bio-Rad
Sourced in Germany
Goat anti-rabbit HRP conjugate is a secondary antibody used in various immunoassay techniques. It contains goat-derived antibodies that specifically recognize and bind to rabbit primary antibodies. The antibodies are conjugated with horseradish peroxidase (HRP), an enzyme that can be used to catalyze a colorimetric or chemiluminescent reaction for detection purposes.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti mouse hrp conjugate, by Bio-Rad (6 mentions)
Hybond, by Cytiva (2 mentions)
Supersignal west femto maximum sensitivity substrate, by Thermo Fisher Scientific (2 mentions)
Mouse anti v5, by Thermo Fisher Scientific (2 mentions)
Rabbit anti gluthathione s transferase, by Thermo Fisher Scientific (2 mentions)
Rabbit anti phospho lrp6 thr1572, by Merck Group (2 mentions)
Goat anti guinea pig hrp conjugate, by Jackson ImmunoResearch (2 mentions)
Blotting grade blocker, by Bio-Rad (1 mentions)
Anti p63, by Abcam (1 mentions)
Anti p27, by BD (1 mentions)
Anti β actin ac 15, by Merck Group (1 mentions)
Amersham hybond p0.45 pvdf membrane, by GE Healthcare (1 mentions)
Anti p63α, by Cell Signaling Technology (1 mentions)
Anti klf4, by R&D Systems (1 mentions)
Anti k10, by BioLegend (1 mentions)
Anti lor, by BioLegend (1 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (1 mentions)
Protease phosphatase inhibitor cocktail, by Cell Signaling Technology (1 mentions)
Ab203383, by Abcam (1 mentions)
Amersham hyperfilm ecl, by GE Healthcare (1 mentions)
12 protocols using goat anti rabbit hrp conjugate
1
Western Blot Analysis of Cellular Proteins
2
Western Blot Analysis of Lung Protein
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Snap-frozen lungs were homogenized on ice in RIPA buffer containing protease/phosphatase inhibitor cocktail (Cell Signaling Technology, Danvers, MA, United States of America). Samples were sonicated and centrifuged at 11000 rpm for 20 min at 4°C. Protein content in the supernatant was quantified using a BCA protein assay kit (Thermo Fisher Scientific, Waltham, MA, United States of America). Twenty micrograms of protein sample per lane were subjected to SDS-PAGE, and proteins from the gel were transferred to PVDF membranes by electroblotting. Immunodetection was performed with a rabbit anti-protein kinase G (PKG) −1 monoclonal antibody (#3248, Cell Signaling Technology) diluted 1:1000 and a rabbit monoclonal to vascular endothelial growth factor A (VEGFA) antibody (ab214424, abcam, Cambridge, UK) diluted 1:1000 overnight at 4°C. After the blots were washed to remove unbound antibody, secondary antibody, Goat anti-rabbit HRP conjugate (1:2000, Bio-Rad, Hercules, CA, United States of America) detection was applied for 1 h at room temperature. After being washed, bands were visualized using an enhanced chemiluminescence kit (SuperSignal West Femto Maximum Sensitivity Substrate, Thermo Fisher Scientific). Additionally, each gel was stripped and reprobed with actin or tubulin as a housekeeping protein to normalize for protein loading.
3
Western Blot Analysis of Myogenic Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Teratomas were homogenized and lysed in RIPA buffer containing protease inhibitors (Complete, Roche), and obtained lysates were kept on ice for 30 min, centrifuged, and stored at − 80 °C. Protein concentration was determined by using Bradford’s assay (Sigma-Aldrich). Twenty microgram protein was subjected to 10% SDS-PAGE and Western blot analysis and probed with antibodies against Pax3 (ARP32446, Aviva; 1:1000), Myf5 (SAB 4501943, Sigma-Aldrich; 1:1000), MyoD (Ab203383, Abcam; 1:1000), Myogenin (sc-576, Santa Cruz Biotechnology; 1:1000), Mck (SAB4500267, Sigma-Aldrich; 1:1000), M-cadherin (SAB4500040, Sigma; 1:1000), or Hsp90 (TA500494, OriGene Technologies). As secondary antibodies, goat anti-rabbit HRP conjugate (170-6515, Bio-Rad) was used, followed by chemiluminescence detection. Films (Amersham Hyperfilm ECL (GE Healthcare)) exposed on membranes were photographed, and optical density of resulting bands was measured using GelDocXR+ (Bio-Rad) with ImageLab software.
4
Protein Immunoblot Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples were separated by Sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to PVDF membranes. Blots were blocked in 5% milk in Tris-buffered saline with Tween 20. Membranes were probed with an anti-GFP (Novus), anti-FLAG (Thermo Fisher Scientific), or anti-HA (Millipore Sigma) primary antibody and goat anti-rabbit HRP conjugate (BioRad) secondary antibody. To evaluate secretion, anti-GSK or anti-P-GSK (Cell Signaling) antibodies were used. Results were collected from at least three independent experiments.
5
Co-Immunoprecipitation of Circadian Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein cross linking was performed using 2mM DSP (D3669; Sigma) using a protocol previously described [25 (link)]. Co-IP was performed as follows: 1mg of total protein incubated with anti-V5 antibody-coated agarose beads (A7345; Sigma) overnight at 4°C followed by six washes with cold protein extraction buffer. Elution was performed by incubating the beads with 2x Western blot sample buffer at 65°C for 20 minutes. Samples to detect proteins were run on pre-made gels (Novex, Life Sciences) and transferred onto PVDF membranes. Polyclonal antibodies were used for FRQ (1:250), WC-1 (1:250) and WC-2 (1:5000) and commercial monoclonal V5 antibody (1:5000, Invitrogen) was used for VVD. Goat Anti-Rabbit HRP conjugate and Goat Anti-Mouse HRP conjugate (BioRad) were used a secondary antibodies. Membranes for anti-V5 IP were developed using SuperSignal West Femto Maximum Sensitivity Substrate (Thermo Scientific) and the signal was captured using X-ray film (GE Healthcare). Western Blots were quantified using NIH ImageJ software.
6
Western Blot Analysis of E. coli and Y. pseudotuberculosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell pellets of E. coli DH5α, Y. pseudotuberculosis YPIII, Y. pseudotuberculosis YP216 were resuspended in 1 x SDS sample buffer (2% (w/v) SDS, 0.1% (w/v) bromophenol blue, 10% glycerol, 50 mM Tris/HCl, pH 6.8) according to their optical density (CNFY synthesis studies: 100 μl for OD600 = 1; gfp reporter gene assay: 100 μl for OD600 = 0.5). After boiling for 10 min at 95°C, samples were centrifugated (10 min, 13000 rpm) and the protein extracts were subjected to SDS gel electrophoresis (10–12.5% SDS PAA gels). Subsequent Western transfer was performed by tank blotting onto a nitrocellulose membrane (Hybond-C Extra, GE Healthcare, Munich, Germany). An anti-CNFY antibody was used in a 1:1000 dilution, anti-GFP antibody (ABIN129570, antibodies-online GmbH, Aachen) was used in a 1:10000 dilution and secondary antibody goat anti-rabbit-HRP conjugate (Bio-Rad, Munich, Germany) in a 1:3000 dilution. Luminescence signals were detected by incubating membranes with Luminata Forte Western HRP (Merck, Darmstadt, Germany) substrate and the ChemiImager Ready (Alpha Innotec, San Leandro, USA).
7
Antibody Immunoblotting and Immunostaining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The primary antibodies used for immunoblotting were mouse anti-V5 (1:5000, Invitrogen), guinea pig anti-Axin (1:1000, [17 (link)]), rabbit anti-Kinesin Heavy Chain (1:10000, Cytoskeleton), mouse anti-alpha-Tubulin (1:10000, DM1A, Sigma), rabbit anti-alpha-Tubulin (1:10000, Sigma), rabbit anti-Gluthathione-S-Transferase (1:10000, Invitrogen), rabbit anti-phospho-LRP6 [Thr1572] (1:1000, Millipore), mouse anti-Nervana antibody (Nrv5F7, 1:1000, DSHB), and guinea pig anti-Arrow antibody (1:1000, [74 (link)]). The primary antibodies used for immunostaining were guinea pig anti-Axin (1:1000, [17 (link)]), rabbit anti-β-gal (1:1000; MP Biomedicals), mouse anti-Arm (1:20; DSHB), mouse anti-Fas III (1:20; DSHB), mouse anti-Discs Large (1:20; DSHB), rabbit anti-GFP (1:200; Invitrogen), mouse anti-V5 (1:5000; Invitrogen), rabbit anti-Apc2 (1:1000; [75 (link)]), and guinea pig anti-Senseless (1:1000, [76 (link)]).
The secondary antibodies used for immunoblotting were: goat anti-rabbit HRP conjugate (1:10000, Biorad), goat anti-mouse HRP conjugate (1:10000, Biorad), and goat anti-guinea pig HRP conjugate (1:10000, Jackson ImmunoResearch). The secondary antibodies used for immunostaining were goat or donkey Alexa Fluor 488, 555 or Cy5 conjugates (1:400; Invitrogen).
The secondary antibodies used for immunoblotting were: goat anti-rabbit HRP conjugate (1:10000, Biorad), goat anti-mouse HRP conjugate (1:10000, Biorad), and goat anti-guinea pig HRP conjugate (1:10000, Jackson ImmunoResearch). The secondary antibodies used for immunostaining were goat or donkey Alexa Fluor 488, 555 or Cy5 conjugates (1:400; Invitrogen).
8
Western Blot Analysis of Hippocampal Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For Western blots, 40 μg of protein extracted from the mouse hippocampal tissue was mixed with 5× SDS-PAGE loading dye and boiled at 95°C for 5 min in a shaking incubator (KTM-100, KBT, Seongnam, Korea). Proteins were then run at 80 V for 20 min across a 14% polyacrylamide gel, followed by 120 V for 1 hr at RT. Protein in the gel was transferred to a methanol-activated PVDF membrane at 400 mA for 40 min on ice. The membrane was blocked in 5% bovine serum albumin (BSA) (160069, MP Biomedicals, Santa Ana, CA) diluted in 1x PBS for 1 hr at RT, and then incubated with primary antibodies (Bax rabbit mAb, 14796, Cell Signaling Technology, Danvers, MA; Recombinant TNF alpha antibody, ab183218, Abcam, Cambridge, UK; β-tubulin mouse mAb, 86298, Cell Signaling Technology) diluted in 2.5% BSA solution with 0.2% Tween-20 at 4°C overnight. The next day, the membrane was incubated with secondary antibodies (goat anti-rabbit HRP conjugate, 170-6515, Bio-Rad, Hercules, CA; goat anti-mouse HRP conjugate, STAR207P, Bio-Rad). The conjugates were activated with enhanced chemiluminescence substrates (#1705061, Bio-Rad) and visualized under a LuminoGraph chemiluminescent imaging system (WSE-6200H, ATTO, Daejeon, Korea).
9
Lentiviral Vector-Based Transfection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Vectors pLenti6/V5-DEST (VT1476), pSPAX2 (VT1444), pMD2.G (VT1443), PET28a, and pCMV-HA (VT1710) were acquired from Youbio. The Luc-IFN-β and pTK-RL were bought from Yeasen. Poly (I:C) was procured from MERCK (P1530). Genes encoding non-structural proteins were previously constructed and preserved by our laboratory.
The following antibodies were used for IFA and IHC: Mouse anti-SR30 (Rtilab, SR30-A), Goat Anti-Mouse HRP (Bioss, bs-0368G-HRP), Goat Anti-Mouse FITC (Bioss, bs-0368G-FITC).
For protein immunoblotting: Rabbit anti-GATM antibody (HPA026077, Sigma), Rabbit anti-NMRAL1 antibody (K004351P, Solarbio), Mouse anti-HA antibody (26D11, Abmart), Mouse anti-GAPDH (60004, Proteintech), Rabbit anti-PRRSV-N (bs-41387R, Bioss), Goat Anti-Mouse HRP Conjugate (1705047, Bio-rad), Goat anti-Rabbit HRP Conjugate (1705046, Bio-rad).
The following antibodies were used for IFA and IHC: Mouse anti-SR30 (Rtilab, SR30-A), Goat Anti-Mouse HRP (Bioss, bs-0368G-HRP), Goat Anti-Mouse FITC (Bioss, bs-0368G-FITC).
For protein immunoblotting: Rabbit anti-GATM antibody (HPA026077, Sigma), Rabbit anti-NMRAL1 antibody (K004351P, Solarbio), Mouse anti-HA antibody (26D11, Abmart), Mouse anti-GAPDH (60004, Proteintech), Rabbit anti-PRRSV-N (bs-41387R, Bioss), Goat Anti-Mouse HRP Conjugate (1705047, Bio-rad), Goat anti-Rabbit HRP Conjugate (1705046, Bio-rad).
10
Antibody Immunostaining and Immunoblotting Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The primary antibodies used for immunostaining were mouse anti-V5 (1:5000; Invitrogen), mouse anti-Wingless (1:200, 4D4 concentrated antibody, Developmental Studies Hybridoma Bank, DSHB). The secondary antibodies used for immunostaining were goat or donkey Alexa Fluor 488 or 555 conjugates (1:400; Invitrogen). The primary antibodies used for immunoblotting were mouse anti-V5 (1:5000, Invitrogen), guinea pig anti-Axin (1:1000, [54 (link)]), rabbit anti-Kinesin Heavy Chain (1:10000, Cytoskeleton), mouse anti-Arm (1:100, N2 7A1, DSHB), mouse anti-alpha-Tubulin (1:10000, DM1A, Sigma), rabbit anti-alpha-Tubulin (1:10000, Sigma), rabbit anti-Gluthathione-S-Transferase (1:10000, Invitrogen), guinea pig anti-Apc2 (GP10) 1:5000 [44 (link)]; rabbit anti-phospho-LRP6 [Thr1572] (1:1000, Millipore), rat anti-Dishevelled (1:1000, [70 (link)]), and guinea pig anti-Arrow (1:1000, [71 (link)]). The secondary antibodies used for immunoblotting were: goat anti-rabbit HRP conjugate (1:10000, Biorad), goat anti-mouse HRP conjugate (1:10000, Biorad), and goat anti-guinea pig HRP conjugate (1:10000, Jackson ImmunoResearch).
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!