Immunoblotting analysis of the Ats-1 protein was performed through SDS-PAGE (CWBIO, Beijing, China) using mouse anti-6 × His-tag antibody (Abcam, Cambridge, MA, USA) (1:1000) as the primary antibody. Goat Anti-mouse IgG H&L (Abcam, Cambridge, MA, USA) (1:3000) was implemented as the secondary antibody, and a SuperSignal West Femto Trial Kit (ThermoFisher, Waltham, MA, USA) was utilized for color development. Actin was utilized as a control. Mouse Anti-β-actin Monoclonal Antibody (Sino Biological, Beijing, China) (1:1000) was used as the primary antibody and Goat Anti-mouse IgG H&L (Abcam, Cambridge, MA, USA) (1:3000) was used as the secondary antibody in our research. And the blots were cut prior to hybridisation with antibodies during blotting.
Goat anti mouse igg h l
Manufactured by Abcam
Sourced in United States, United Kingdom, China
Goat anti-mouse IgG H&L is a secondary antibody used in various immunodetection techniques. It is raised in goat against mouse immunoglobulin G (IgG) heavy and light chains. This antibody can bind to mouse IgG, allowing for the detection and visualization of mouse primary antibodies in applications such as Western blotting, ELISA, and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti rabbit igg h l, by Abcam (33 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (5 mentions)
Supersignal west femto trial kit, by Thermo Fisher Scientific (4 mentions)
Ab205719, by Abcam (4 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (3 mentions)
Ab6721, by Abcam (3 mentions)
Ab6789, by Abcam (3 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (3 mentions)
Ripa lysis buffer, by Beyotime (3 mentions)
Fluorescence microscope, by Olympus (3 mentions)
Mouse anti β actin monoclonal antibody, by Sino Biological (2 mentions)
Sds page, by CWBIO (2 mentions)
Mouse typer isotyping panel, by Bio-Rad (2 mentions)
Alexa fluor 488, by Abcam (2 mentions)
Immunol staining blocking buffer, by Beyotime (2 mentions)
4 compartment cell view cell culture dishes, by Greiner (2 mentions)
Ab96879, by Abcam (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
β actin, by Santa Cruz Biotechnology (2 mentions)
Pvdf membrane, by Merck Group (2 mentions)
97 protocols using goat anti mouse igg h l
1
Immunoblotting Analysis of Ats-1 Protein
2
Fluorescent Immunocytochemical Staining Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were fixed with 4 % paraformaldehyde (PFA) at 4 °C for 48 h, rinsed three times with PBS, then incubated with TritonX-100 (Solarbio, P1080) at room temperature for 20 min to permeabilize cells. Block non-specific reaction with 10 % FBS PBS at room temperature for 1 h, and resuspend primary antibody in 5 % BSA PBS overnight at 4 °C. Cells were rinsed three times and incubated with the appropriate fluorochrome-conjugated secondary antibodies and DAPI (ab285390) at room temperature for 1 h, then washed out antibodies and DAPI with PBS for three times before taking images using an Axio Observer 3 (Zeiss)microscopy [21 ]. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488, ab150077), Goat Anti-Mouse IgG H&L (Alexa Fluor® 488, ab150113), Goat Anti-Mouse IgG H&L (Alexa Fluor® 647, ab150115) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647, ab150079) were purchased from the Abcam.
3
Antibody Characterization for Alzheimer's Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used in this study: anti-Fyn (catalogue no. ab1881, Abcam), anti-Fyn (catalog no.: sc-434, Santa Cruz Biotechnology), anti-APP (catalog no.: ab32136, Abcam), anti-actin (catalog no.: TA-09, Zsgb-Bio, China), anti-NMDAR2B (catalog no.: ab254356, Abcam), anti-ZDHHC21 (catalog no.: PA5-25,096, Invitrogen, USA), anti-NeuN (catalog no.: ab279295, Abcam), anti-tau (phospho T231; catalog no.: ab151559, Abcam), anti-tau (phospho Thr181; catalog no.: 701530, Invitrogen), anti-Aβ (catalog no.: ab201060, Abcam), anti-phospho-Src family (catalog no.: 2101S, Cell Signaling Technology, USA), horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG heavy chain-specific secondary antibody (catalog no.: A25222, Abbkine, China), HRP-conjugated mouse anti-rabbit IgG light chain (catalog no.: A25022, Abbkine), goat anti-mouse IgG H&L (HRP; catalog no.: ab6789, Abcam), goat anti-rabbit IgG H&L (HRP; catalog no.: ab205718, Abcam), goat anti-rabbit IgG H&L (Cy3; catalog no.: ab6939, Abcam), and goat anti-mouse IgG H&L (FITC; catalog no.: ab6785, Abcam).
4
Immunofluorescence and Western Blotting of Neuronal and ER Stress Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The primary antibodies include anti-MAP-2 (1:1,000, Abcam, catalog no. ab32454) anti-GABAAR a1 (1:1,000, Abcam, catalog no. ab94585), anti-BiP (1:1,000, Abcam, catalog no. ab21685), anti-CHOP (1:500 for immunofluorescence, 1:1,000 for Western blotting, Cell Signaling Technology, catalog no. 2895), anti-β-actin (1:1,000, Abcam, catalog no. ab8226), anti-pan-cadherin (1:1,000, Abcam, catalog no. ab16505), anti-β-tubulin (1:1,000, Abcam, catalog no. ab18207), and anti-ubiquitin (1:1,000, Abcam, catalog no. ab134953). The secondary antibodies include Goat Anti-Mouse IgG H&L [horseradish peroxidase (HRP)] (1:2,000, Abcam, catalog no. ab205718), Goat Anti-Mouse IgG H&L (HRP; 1:2,000, Abcam, catalog no. ab250719), Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488; 1:1,000 for immunofluorescence, ab150077), and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594; 1:1,000 for immunofluorescence, ab150120).
5
Protein Expression Analysis in HUVECs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total protein was extracted from HUVECs using radioimmunoprecipitation assay buffer containing 1 mM PMSF. Bicinchoninic Acid Protein assay kit (Thermo Fisher Scientific, Inc.) was then used to quantify protein concentration following which the protein samples were separated by 10% SDS-PAGE (20 µg/lane) and transferred onto PVDF membranes. Subsequently, the membrane was blocked with 5% skim milk for 1 h at room temperature and incubated with primary antibodies against phosphorylated (p)-AKT (cat. no. ab81283; 1:1,000; Abcam), AKT (cat. no. ab179463, 1:10,000, Abcam), p-PI3K (cat. no. ab182651, 1:1,000, Abcam), PI3K (cat. no. ab86714, 1:1,000, Abcam), eNOS (cat. no. ab66127, 1:1,000, Abcam) or GAPDH (cat. no. ab181602, 1:10,000, Abcam) at 4°C overnight. The membrane was then incubated with horseradish peroxidase-conjugated secondary antibody Goat Anti-Rabbit IgG H&L (ab205718; 1:2,000; Abcam), Goat Anti-Mouse IgG H&L (ab205719; 1:2,000; Abcam) and for 2 h at room temperature. The GAPDH was used as internal reference. Afterwards, the protein bands were detected and visualized using an enhanced chemiluminescence method (RapidStep™ ECL Reagent; Merck KGaA). The densities of bands were semi-quantitively analyzed using ImageJ (version 4.0; National Institutes of Health).
6
Quantification of IgG Subclasses by EIA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An EIA was performed by sorbing on immune plates of goat anti-mouse IgG (Goat Anti-Mouse IgG H&L, 6708, Abcam, USA), followed by the titration of immune and nonimmune sera. After incubation and washing, rabbit antibodies against IgG subclasses (Bio-Rad, mouse typer isotyping panel, 1722055, USA) were added. Then, the mixture was incubated for 1 h and washed. Next, a conjugate of goat antibodies against rabbit immunoglobulins with peroxidase (Imteck, 110810, Russia) was added as recommended by the manufacturer. Staining was also carried out with an ortho-phenylenediamine solution. Wells without sera were used as negative-controls. Monoclonal antibodies of the IgG1, IgG2a, and IgG2b subclasses that were previously identified by this kit were used as calibrators and made into serial dilutions with steps of 2 starting at a concentration of 1 μg · mL−1 in wells with immobilized goat anti-mouse antibodies.
The determination of K9-binding IgG subclasses was performed similarly, but the K9 were immobilized on the immunoplate’s wells from a 1 μg · mL−1 solution. The serum and monoclonal antibodies’ preparations were measured in three repeats, each in a series of eight dilutions, starting from 1/100 for the serum of each mouse, separately. The contents of the IgG subclasses were calculated as described in (34 (link), 52 (link)).
The determination of K9-binding IgG subclasses was performed similarly, but the K9 were immobilized on the immunoplate’s wells from a 1 μg · mL−1 solution. The serum and monoclonal antibodies’ preparations were measured in three repeats, each in a series of eight dilutions, starting from 1/100 for the serum of each mouse, separately. The contents of the IgG subclasses were calculated as described in (34 (link), 52 (link)).
7
Immunofluorescence Staining of iPSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fixed iPSCs were permeabilized with 0.05% Triton X-100 for 30 minutes at room temperature (RT). Cells were incubated with primary antibodies OCT4 and SSEA4 (1:100, Stemgent) during an overnight at 4°C. Cells were washed twice and incubated with secondary goat anti-mouse IgG H&L (1:700, Alexa Fluor 488, Abcam), or goat anti-rabbit IgG H&L (1:700, Alexa Fluor 555, Abcam) antibodies for 1 h at RT.
8
Western Blot Analysis of NUCB-2 Protein
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed using radioimmunoprecipitation assay buffer (EMD Millipore) on ice for 30 min. The total cell lysate was collected after centrifugation at 12,000 × g for 15 min at 4°C. For extraction of nuclear proteins, a Nuclear Extract kit (Active Motif) was used. The concentration of protein was determined using a bicinchoninic acid protein assay kit (EMD Millipore). Proteins (50 µg) were separated by 6–12% SDS-PAGE and transferred onto polyvinylidene difluoride membranes. After transfer, 5% non-fat dry milk was used to block the membranes for 1 h. The membranes were incubated with rabbit anti-human NUCB-2 polyclonal antibody (cat. no. ab224348, 1:100, Abcam) and recombinant GAPDH antibody (cat. no. ab181602, 1:100, Abcam) at 4°C for overnight. Membranes were washed three times in TBST for 15 min and incubated with an HRP goat anti-rabbit IgG H&L (cat. no. ab205718; 1:200; Abcam) and goat anti-mouse IgG H&L (cat. no. ab150117; 1:200; Abcam) at room temperature for 1 h. Signals were visualized using enhanced chemiluminescence substrate (EMD Millipore) on an imaging capture system (Alpha Imaging).
9
Western Blotting Assay Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting assays were performed according to methods described previously (Hua et al., 2022 (link)). The following antibodies were utilized for Western blotting: Bam (mouse, 1:3000, DSHB), Actin (mouse, 1:5000, Cat#A4700; Sigma), and Goat anti-Mouse IgG H&L (1:3000, Cat#ab150078; Abcam).
10
Protein Extraction from Trophozoite-Infected Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 5×105 RAW264.7 or CHO-K1 cells were cultured in a 24-well plate. Once the cells completely adhered to the slide, 5×105 trophozoites were added to the wells. After incubating at 36.5°C for 1 h in relatively anaerobic environment, the culture supernatant was collected. Then, cold acetone precipitation method was applied to extract protein from the culture supernatant. The extracted protein was resuspended in 30 μL loading buffer (containing β-ME). Trophozoite lysate was used as the positive control. The protein samples were separated by 10% SDS-PAGE and detected by western blotting. 4G6 (mouse ascites, 1:400), and Goat anti mouse IgG (H + L) (Abcam, UK) were used as the primary and secondary antibodies, respectively.
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!