The mAbs used for flow cytometric analysis and other experiments were obtained from eBioscience (San Diego, CA, USA) or BD Biosciences (San Diego, CA, USA): FITC-labeled anti-mouse CD4 (RM4-5), CD45 (30-F11); PE-labeled anti-mouse IFN-γ (XMG1.2), FoxP3 (FJK-16s); PerCP-labeled anti-mouse CD4 (L3T4, RM4-5); allophycocyanin (APC)-conjugated anti-mouse IL-17A (eBio17B7); biotin-conjugated anti-mouse IL-4 (BVD6-24G2), and streptavidin-conjugated PerCP. PMA and ionomycin were purchased from Sigma-Aldrich (St. Louis, MO, USA). The Abs used to detect OVA-specific-IgE, IgG, IgG1, and IgG2a levels in sera were purchased from Southern Biotech (Birmingham, AL, USA): unconjugated (human absorbed) anti-mouse IgG, goat anti-mouse IgG1, goat anti-mouse IgG2a, goat anti-mouse IgE, HRP-conjugated goat anti-mouse IgG, goat anti-mouse IgG1, goat anti-mouse IgG2a, and rat anti-mouse IgE-HRP. The primers specific for cytokines, mucins, and tight junctions (TJs) were synthesized by Bioneer Corp. (Daejeon, Korea) and used for PCR amplification of target genes.
Goat anti mouse igg1
Manufactured by Southern Biotech
Sourced in United States, United Kingdom
Goat anti-mouse IgG1 is a secondary antibody produced in goats that specifically binds to the IgG1 subclass of mouse immunoglobulins. This antibody can be used in various immunodetection techniques, such as Western blotting, ELISA, and immunohistochemistry, to identify and quantify mouse IgG1 proteins.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti mouse igg2a, by Southern Biotech (8 mentions)
Goat anti mouse igg2b, by Southern Biotech (5 mentions)
Enhanced chemiluminescence, by Cytiva (3 mentions)
Goat anti mouse igg, by Santa Cruz Biotechnology (3 mentions)
Goat anti rabbit igg conjugated to horseradish peroxidase, by Santa Cruz Biotechnology (3 mentions)
Donkey anti goat igg, by Santa Cruz Biotechnology (3 mentions)
Mouse anti β actin, by Merck Group (3 mentions)
Goat anti mouse iga, by Southern Biotech (3 mentions)
Goat anti mouse kappa antibodies, by Southern Biotech (2 mentions)
Mouse anti akt, by BD (2 mentions)
Rabbit anti p 4e bp1, by Cell Signaling Technology (2 mentions)
Rabbit anti 4e bp1, by Cell Signaling Technology (2 mentions)
Rabbit anti ps6, by Cell Signaling Technology (2 mentions)
Mouse anti s6, by Cell Signaling Technology (2 mentions)
Mouse anti gapdh, by HyTest (2 mentions)
Goat anti mouse ig, by Southern Biotech (2 mentions)
Mouse anti caspase 9, by Cell Signaling Technology (2 mentions)
Mouse anti caspase 8, by Alexis Biochemicals (2 mentions)
Rabbit anti goat igg conjugated with horseradish peroxidase, by Southern Biotech (2 mentions)
Goat anti mouse igg conjugated with horseradish peroxidase, by Merck Group (2 mentions)
33 protocols using goat anti mouse igg1
1
Antibody-based Immunological Analyses
2
ELISA for Virus-Specific Antibody Titers
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Purified β-propiolactone-inactivated rWT virus (2.5 μg/mL) was coated onto 96-well Immulon-2 plates (Dynex Technology Inc., Chantilly, VA, USA) and incubated overnight at 4°C. Serially diluted mouse sera was added and left to incubate at room temperature for 1.5 h. Goat anti-mouse IgG (H+L) (Invitrogen, B2763, Carlsbad, CA, USA), goat anti-mouse IgG1 (Southern Biotech, 1070-08, Birmingham, AL, USA), or goat anti-mouse IgG2a (Southern Biotech, 1080-08, Birmingham, AL, USA) was added for 1 h at room temperature to detect IgG, IgG1, or IgG2a, respectively. Color development was initiated by the addition of alkaline phosphatase (AP)-conjugated streptavidin (Jackson ImmunoResearch) and p-nitrophenyl phosphate (PNPP) substrate [10 mg/mL p-nitrophenyl phosphate di (tris) salt crystalline (Sigma-Aldrich, St. Louis, MO, USA), 1% diethanolamine (Sigma-Aldrich, St. Louis, MO, USA), 0.5 mg/mL MgCl2, pH 9.8]. Detection of the optical density (OD) occurred at 405 nm (using a reference filter of 490 nm) on a microplate reader (Molecular Devices SpectraMax Plus 384, San Jose, CA, USA). The titres of each sample were determined as the highest dilution at which the OD of the sample was larger than the defined cut-off. The cut-off was defined as the mean OD of a known negative sample plus twice the standard deviation.
3
Quantifying Serum Clearance of 1C6 mAb and Fab
4
Western Blot Analysis of Protein Signaling
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Western blot analysis was performed as described previously (12 (link)) using the following antibodies: mouse anti-AKT (BD Biosciences), rabbit anti-pAKT, rabbit anti-p4E-BP1, rabbit anti-4E-BP1, rabbit anti-pS6, mouse anti-S6, rabbit anti-pERK, rabbit anti-ERK, rabbit anti-pan-RAS (Cell Signaling, Beverly, MA, USA). Mouse anti-GAPDH (HyTest, Turku, Finland) or mouse anti-β-Actin (Sigma) were used as loading controls. Goat anti-mouse IgG, donkey anti-goat IgG, goat anti-rabbit IgG conjugated to horseradish peroxidase (Santa Cruz Biotechnology Inc.), and Goat anti-mouse IgG1 or Goat anti-mouse IgG2b (Southern Biotech, Birmingham, AL, USA) conjugated to horseradish peroxidase were used as secondary antibodies. Enhanced chemiluminescence was used for detection (Amersham Bioscience, Freiburg, Germany). Also, donkey anti-mouse IgG or donkey anti-rabbit (LI-COR Biotechnology, Bad Homburg, Germany) labeled with IRDye infrared dyes were used for detection. Representative blots of at least two independent experiments are shown.
5
Bat Serum IgG Assay Development
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Sera from several distinct Megachiropteran bat species were purchased through cooperation with the Lubee Bat Conservancy (Gainesville, FL). Purified IgG derived from dog, cat, guinea pig, swine, hamster, human were purchased (Rockland Immunochemicals). Laboratory stocks of mouse and rat mAbs were used as unlabeled IgG or conjugated with EZ-Link Sulfo-NHS-LC-LC-Biotin (Thermo Scientific); horse serum was obtained from the Wadsworth Center Veterinary Sciences facility. For ELISAs, unlabeled and horseradish peroxidase (HRP)- conjugated goat anti-mouse Ig, goat anti-mouse IgG1, goat anti-mouse IgG2a, and goat anti-mouse IgM, as well as HRP-streptavidin were used (Southern Biotech). In addition, polyclonal HRP-conjugated goat anti-bat Ig was purchased (Bethyl Laboratories). For flow cytometry, rabbit anti-mouse Ig (RamIg) (Jackson Research, West Grove, PA) was purchased and labeled with FITC (Sigma-Aldrich) or Cy-5 (GE HealthCare).
6
IgE and IgG1 Measurement for HDM Allergy
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Total IgE was measured in the serum by using a mouse IgE ELISA set (BD Biosciences) according to supplier’s recommendations. An indirect ELISA method was used to measure HDM-specific IgG1 levels in serum samples as previously described by Trompette et al. (22 (link)). Briefly, 96-well microtiter plates were coated overnight with 100 µl of HDM at 10 µg/ml in PBS. The next day, 200 µl of blocking solution (1% BSA in PBS) was added to the plate for 2 h at room temperature. Subsequently, 100 µl of serum sample diluted 1:100 and 1:500 in blocking buffer was added to the plate at 4°C overnight followed by goat-anti-mouse IgG1 (Southern Biotech) for 1 h. Then HRP-donkey anti-goat IgG (Santa Cruz) was added to the plate for 1 h at room temperature, followed by the substract TMB. Absorbance was measured at 450 nm using a microplate reader (VersaMax microplate reader, Molecular Devices). No HDM-specific IgG1 was detected in control mice.
7
Antibody Response Evaluation by ELISA
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Seven days after last immunisation, the sera of anti-rMntC and anti-immunodominant peptide were collected from the tail vein for analysis of IgG and other antibodies using ELISA. The assay of antibody detection by using ELISA was as described previously[21 (link)]. The corresponding collected serum samples from mice immunised with rMntC and epitope vaccination were employed as the primary antibodies and the appropriate horseradish peroxidase-conjugated goat anti-mouse IgG (Dianova, Hamburg, Germany), or goat anti-mouse IgG1, or goat anti-mouse IgG2a, or goat anti-mouse IgG2b (Southern Biotech, Birmingham, AL, USA) were used as the secondary antibody. The highest absorbance was calculated by log test from the reduplicate assay and blank control and the comparison between the two results was undertaken by t-test.
8
Western Blot Analysis of Apoptosis Regulators
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Western blot analysis was performed as described previously [52 (link)] using the following antibodies: mouse anti-caspase-8, mouse anti-NOXA, rat anti-BMF (Alexis Biochemicals, Grünberg, Germany), mouse anti-AKT, mouse anti-BCL-2, mouse anti-BAX, rabbit anti-BAK (BD Bioscience), rabbit anti-caspase-3, mouse anti-caspase-9, rabbit anti-pAKT, rabbit anti-p4E-BP1, rabbit anti-4E-BP1, rabbit anti-pS6, mouse anti-S6, (Cell Signaling, Beverly, MA), rabbit anti-MCL-1 (Stressgene Bioreagents, Victoria, BC). Mouse anti-GAPDH (HyTest, Turku, Finland) or mouse anti-β-Actin (Sigma) were used as loading controls. Goat anti-mouse IgG, donkey anti-goat IgG, goat anti-rabbit IgG conjugated to horseradish peroxidase (Santa Cruz Biotechnology, Santa Cruz, CA)) and Goat anti-mouse IgG1 or Goat anti-mouse IgG2b (Southern Biotech, Birmingham, AL) conjugated to horseradish peroxidase were used as secondary antibodies. Enhanced chemiluminescence was used for detection (Amersham Bioscience, Freiburg, Germany). Representative blots of at least two independent experiments are shown.
9
ELISA Protocol for PspA4Pro Antibody
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ELISA was carried out as described previously [12 (link)] in plates coated with 5 μg/ml PspA4Pro. For the detection of serum antibodies, goat anti-mouse IgG conjugated with horseradish peroxidase (Sigma-Aldrich) was used as secondary antibody. For IgG isotyping, goat anti-mouse IgG1, goat-anti-mouse IgG2a and rabbit anti-goat IgG conjugated with horseradish peroxidase (Southern Biotech) were used. The titer was defined as the reciprocal of the highest dilution with an A492 ≥ 0.1. For the detection of antibodies in BALF, goat anti-mouse IgG conjugated with alkaline phosphatase (AP), goat anti-mouse IgA and rabbit anti-goat IgG conjugated with AP (Southern Biotech) were used and A405 was measured. Background value of each plate (0.040 to 0.045) was subtracted from the actual absorbance obtained for each sample.
10
ELISA Assay for OVA-Specific Antibodies
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OVA-specific antibodies in serum samples were measured by IgG total or isotype specific ELISA. The plates were coated overnight at 4°C with OVA (2 µg/ml in 0.05 M carbonate buffer, pH 8.2) and blocked with 3% w/v bovine serum albumin (BSA) in PBS for 1 h at 37°C. Serial 2-fold dilutions of the samples were prepared in 3% w/v BSA in PBS and incubated for 2 h at 37°C, washed 6 times with 0.1% v/v Tween20 in PBS. The biotinylated detection antibodies goat, anti-mouse IgG (Sigma-Aldrich), goat anti-mouse IgG1 or goat, anti-mouse IgG2c (Southern biotech, USA), 1∶5000 in 1% w/v BSA/0.1% v/v Tween20 in PBS, were used for incubation for 2 h at 37°C. Six washes were followed by incubation with Streptavidin-horse radish peroxidase (HRP) (BD Pharmingen), 1∶1000 in 1% w/v BSA/0.1% v/v Tween20 in PBS, for 1 h at 37°C, six washes, incubation with ABTS [2,20-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)] in 0.1 M citrate-phosphate buffer (pH 4.35) and 0.03% H2O2 (Sigma-Aldrich), and the absorbance of light at 405 nm wave length was measured using a Synergy 2 Multi-Mode Microplate Reader (Biotek, USA). Titers were determined as the highest dilution factors of the assay samples with twice the average readout value of the blank.
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