Rabbit anti-acetyl α-tubulin (K40, #5335), rabbit anti-SIRT2 (#12672) and mouse anti-α tubulin (#3873) were obtained from Cell Signaling Technology (Danvers, MA). Mouse anti-NQO1 (A180, NB200-209) was obtained from Novus Biologicals (Centennial, CO) and rabbit anti-NQO1 (C-terminus, N5288) and mouse anti-β actin (#A2228) were purchased from Sigma-Aldrich. Mouse anti-SIRT2 (#66410-1-lg) was obtained from Proteintech (Rosemont, IL) and used in studies with 3T3-L1 fibroblasts. Horseradish peroxidase-, Alexa fluor 488- or rhodamine Red-X conjugated goat anti-mouse IgG or goat anti-rabbit IgG secondary antibodies were purchased from Jackson ImmunoResearch Laboratories (West Grove, PA).
Goat anti rabbit igg secondary antibody
Manufactured by Jackson ImmunoResearch
Sourced in United States
Goat anti-rabbit IgG secondary antibody is a laboratory reagent used in immunoassays and related techniques. It is a polyclonal antibody produced in goats and specifically binds to rabbit immunoglobulin G (IgG) antibodies. This secondary antibody can be used to detect and amplify signals from primary rabbit antibodies in various applications such as Western blotting, ELISA, and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti mouse igg secondary antibody, by Jackson ImmunoResearch (4 mentions)
Mouse anti gfp primary antibody, by Thermo Fisher Scientific (2 mentions)
Rabbit anti tom20, by Abcam (2 mentions)
Bca protein assay, by Thermo Fisher Scientific (2 mentions)
Ab178870, by Abcam (1 mentions)
Hoechst, by Thermo Fisher Scientific (1 mentions)
Anti nr6a1, by Proteintech (1 mentions)
Gapdh, by Proteintech (1 mentions)
Goat anti mouse igg conjugated with cy3, by Jackson ImmunoResearch (1 mentions)
Ax70 fluorescence microscope, by Olympus (1 mentions)
Saponin, by Merck Group (1 mentions)
α β tubulin antibody, by Cell Signaling Technology (1 mentions)
Supersignal west pico chemiluminescent substrate, by Thermo Fisher Scientific (1 mentions)
Trans blot turbo transfer system, by Bio-Rad (1 mentions)
Horseradish peroxidase, by Jackson ImmunoResearch (1 mentions)
Ab184247, by Abcam (1 mentions)
Gb11001, by Abcam (1 mentions)
Thunder link kit, by Abcam (1 mentions)
Enhanced chemiluminescence detection kit, by GE Healthcare (1 mentions)
Nbp2 47337, by Novus Biologicals (1 mentions)
17 protocols using goat anti rabbit igg secondary antibody
1
Antibody Reagents for Western Blotting
2
IKKβ Knockdown in LLC Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
LLC cells treated with IKKβ shRNA or control shRNA were plated on coverslips coated by laminin in a 24-well plate. After incubation for 36 hours, the cells were fixed with 4% paraformaldehyde and washed three times in PBS. Cell permeabilization and blocking were performed using 0.3% Triton X-100 PBS containing 10% horse serum for 30 minutes at room temperature. After incubation overnight at 4°C with antibodies against IKKβ (ab178870, Abcam, Cambridge, UK), the cells were stained with goat anti-rabbit IgG secondary antibodies (Jackson ImmunoResearch, West Grove, PA, US) at room temperature for one hour and Hoechst (Invitrogen, Carlsbad, CA, US) for nuclear staining for 15 minutes. The labeled cells were immediately imaged by confocal microscopy.
3
Western Blot Protein Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed by grinding the cell precipitate with 2 × SDS lysate, and the proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes that were blocked with 5% milk. The primary antibodies were rabbit anti-human anti-GAPDH (1:3,000, Bioss, Beijing, China) and anti-NR6A1 (1:1,000, Proteintech, Illinois, USA) antibodies. The secondary antibodies were goat anti-rabbit IgG secondary antibodies (1:5,000, Jackson ImmunoResearch, Pennsylvania, USA). Chemiluminescence gel was used for image development.
4
Western Blot Antibody Validation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot assays were performed as described previously [34] using the following antibody: anti-RPL34 (BBI; 1:1000); anti-c-Myc (CST; 1:1000); Goat anti-rabbit IgG secondary antibodies (Jackson ImmunoResearch; 1:5000); Goat anti-mouse IgG secondary antibodies (Jackson ImmunoResearch; 1:5000); GAPDH (Proteintech; 1:5000).
5
Immunofluorescence Imaging of C. muridarum
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HeLa cell monolayers grown on coverslips infected by C. muridarum transformants were fixed with 4% paraformaldehyde for 45 min at room temperature and permeabilized with 2% saponin for an additional 60 min. After blocking, the cell samples were incubated with mouse anti-Pgp3 or GlgA antibody, rabbit antichlamydial organism antibody, and Hochest (to mark DNA) overnight at 4°C. A goat anti-rabbit IgG secondary antibody conjugated with Alexa Fluor 488 and a goat anti-mouse IgG conjugated with Cy3 (Jackson ImmunoResearch, West Grove, PA) were used to incubate the cell samples at room temperature for 1 h. Immunofluorescence images were acquired using an immunofluorescence scanning microscope (Zeiss, Germany).
6
Chlamydia Infection Visualization Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HeLa 229 monolayers cultured on coverslips were infected with C. muridarum strain Nigg and processed for antibody staining as described previously (19 (link), 20 (link)). Infected cells were fixed with 2% paraformaldehyde for 30 min and permeabilized with 1% saponin (Sigma-Aldrich) for 45 min at room temperature. After blocking, Hoechst staining solution (blue; Sangon Biotech Co., Ltd.) was used to visualize DNA. A genus-specific rabbit antibody against chlamydial EBs and Cy2 (green; Jackson ImmunoResearch Laboratories, Inc., West Grove, PA, USA)-labeled goat anti-rabbit IgG secondary antibody were used to visualize the chlamydial inclusions. Antisera collected from each immunized mouse group and Cy3 (red; Jackson ImmunoResearch Laboratories, Inc.)-conjugated goat anti-mouse IgG secondary antibody were used to visualize the Pgp3 antigen. All images were obtained using an AX-70 fluorescence microscope (Olympus, Melville, NY, USA).
7
Quantitative Immunoblotting of PIKfyve and Tubulin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Approximately 2 × 106 U2OS cells expressing or not PIKfyve or PIKfyveKYA were collected and sonicated to extract total proteins. 100 µg of total proteins was then subjected to electrophoresis in 4–12% Bis-Tris Plus Gels (Novex, NW00125BOX) and transferred onto PVDF membrane using a Trans-Blot Turbo Transfer System (Bio-Rad). After blocking with 5% non-fat milk, PVDF membrane was incubated at 4°C with 1:1000 ratio of PIKFYVE antibody (Invitrogen, PA5-13977) or 1:1000 ratio of α/β-tubulin antibody (Cell Signaling, 2148). Both primary antibodies were probed using goat anti-rabbit IgG secondary antibody conjugated with horseradish peroxidase (Jackson ImmunoResearch, West Grove, PA) and visualized by chemiluminescence (SuperSignal West Pico Chemiluminescent Substrate, Thermo Fisher). Images were obtained using an Odyssey FC Imager (Dual-Mode Imaging System; 2 min integration time).
8
Immunofluorescent Labeling of Organelles in U-2 OS Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
U-2 OS cells were electroporated with plasmids encoding endoplasmic reticulum marker mEm-Sec61B (gift from Michael Davidson; Addgene plasmid # 54249). Samples were processed similarly to samples labelled only for mitochondria 25 (link) with the following changes: the cells were labeled with mouse anti-GFP primary antibody (Invitrogen, A-11120) at 1:100 and rabbit anti-TOM20 (Abcam, ab78547) at 1:500 overnight at 4°C. After washing, the samples were labeled with goat anti-mouse IgG secondary antibody (Jackson ImmunoResearch, 115-005-146) and goat anti-rabbit IgG secondary antibody (Jackson ImmunoResearch, 111-005-144) conjugated with oligonucleotide docking strands (Ext. Data Fig. 1 , D1-1c and D2-1a, respectively) both at a concentration of 1:200 for 1 hr at room temperature.
9
Western Blot Analysis of Mitochondrial Dynamics
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All of the tissues or cells were lysed in RIPA buffer, and their total protein concentrations were determined with a BCA protein assay kit. The total protein was loaded into precast 10% SDS-PAGE gels, and then the target protein was transferred to PVDF membranes. The membrane was blocked in 5% milk for 1 h and washed once with Tris-buffered saline with Tween-20 (TBST). After that, the membranes were incubated at 4 °C overnight with the following primary antibodies: anti-β-actin (Servicebio, GB11001, 1:5000), anti-Drp1 (Abcam, ab184247, 1:2000), anti-Mfn2 (Abcam, ab124773, 1:2000), and anti-Opa1 (Proteintech, 27733-1-AP, 1:2000). The primary antibodies were recycled and the membranes were washed 3 times in TBST for 6 min each. Goat anti-Rabbit IgG secondary antibody (Jackson ImmunoResearch, 103069, 1:5000) was incubated for 1 h at room temperature. After removing the secondary antibody, the membranes were washed 3 times with TBST for 6 min each. Finally, the levels of the relative proteins were detected using an enhanced chemiluminescence reagent. The signals were confirmed with a band intensity analysis using Image J software.
10
DNA-antibody conjugation protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Both the “imager” and the “docking” strands (nucleotide sequence and terminal modifications specified in Table S1 ) were commercially synthesized by Eurofins UK. The two nucleotide designs of the P1 and P3 sequences were obtained from Jungmann et al. (2014) (link). We opted for a direct thiolation to link a 5′ C6 amine of the docking strands and cysteines of the antibody. Respective docking strands were conjugated to either a goat anti-mouse immunoglobulin G (IgG) or a goat anti-rabbit IgG secondary antibody (Jackson ImmunoResearch, PA) or a primary antibody against RyR2, clone C3-33, using a Thunder-Link kit (Innova Biosciences, Cambridge) and spectrophotometric analysis to ensure an oligo: antibody conjugation ratio ≥1:1.
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!