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Goat anti rabbit igg antibody

Manufactured by Merck Group
Sourced in United States, Germany

The Goat anti-rabbit IgG antibody is a laboratory reagent used to detect and quantify rabbit immunoglobulin G (IgG) in various immunoassays and research applications. It is a secondary antibody produced in goats that specifically binds to rabbit IgG molecules, allowing for the identification and measurement of rabbit IgG in biological samples.

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24 protocols using goat anti rabbit igg antibody

1

Dexamethasone Induces Mitochondrial Dysfunction

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Dexamethasone (Dex) was purchased from Taiji Group (Chongqing, China). Dulbecco’s Modified Eagle Medium (DMEM) was purchased from Hyclone (Logan, UT, USA). Fetal bovine serum (FBS), L-glutamine and antibiotics, mouse anti-actin antibody, goat anti-mouse IgG antibody and goat anti-rabbit IgG antibody were purchased from Millipore (Billerica, MA, USA). Mouse anti-HO-1 antibody, mouse anti-CypD antibody, rabbit anti-phospho-p38 antibody and rabbit anti-p38 antibody were purchased from abcam (Cambridge, MA, USA). Mitosox red, MitoTracker green and Lipofectamine® RNAiMAX Transfection Reagent were purchased from Thermo Scientific (Waltham, MA, USA). ON-TARGET plus Mouse ppif siRNA (L-062722) and control siRNA (D-001810) were purchased from Dharmacon Research (CO, USA). Annexin V-FITC/propidium iodide (PI) Apoptosis Detection kit was purchased from KeyGEN BioTECH (Nanjing, China). Other reagents not mentioned here were purchased from Sigma-Aldrich (St. Louis, MO, USA).
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2

Immunoblotting of Per a 2 Protein

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Harvested cells of Per a 2 L. lactis were s dispersed in PBS and disturbed by sonication using a Branson digital sonifier (Branson Ultrasonics, Danbury, CT) for 30 minutes on ice. Samples of cell lysates were separated on a 4–12% polyacrylamide gel using Laemmli’s method. For immunoblotting, the gel was moved to a nitrocellulose membrane and following probed with rabbit anti-rPer a 2 antibody. Subsequent detection was performed using a peroxidase-labeled goat anti-rabbit IgG antibody (10000-fold dilution, Millipore) and 3-amino-9-ethyl carbazole (AEC) as a substrate of enzyme.
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3

Protein Expression Analysis in Cancer Cells

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Protein was extracted from HCT15 and A549 cells using CytoBuster Protein Extraction Reagent (Novagen, Madison, WI, USA) following the manufacturer's instructions. Equal amounts of protein (30 μg) were separated on 10% sodium dodecyl sulfate (SDS)-polyacrylamide gels and transferred to a 0.2 μm nitrocellulose membrane (Whatman GmbH, Hahnestraße, Germany). The membranes were blocked by incubating with Tris-buffered saline-Tween 20 (TBST) containing 10% skim milk (Difco, Sparks, MD, USA) for 2 h at room temperature. The blocked membranes were probed with anti-cyclin D3 (1:2000, 2936; Cell Signaling Technology, Danvers, MA, USA), anti-CDK2 (1:1000, sc-163; Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-p21 (1:1000, sc-397; Santa Cruz Biotechnology) or anti-p27 (1:500, sc-528; Santa Cruz Biotechnology) antibody at 4°C overnight. Membranes were incubated with horseradish peroxidase-conjugated horse anti-mouse IgG antibody (Vector Laboratories Inc., Burlingame, CA, USA) or goat anti-rabbit IgG antibody (Millipore, Billerica, MA, USA) for 1.5 h at room temperature. β-actin (Cell Signaling Technology) was used as an internal control.
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4

Autophagy Regulation in DMEM/F12 Cell Culture

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DMEM/F12 (D0697; Thermo Fisher), Fetal bovine serum (FBS) (10099141; Gibco), Penicillin (ST488-1; Beyotime), Streptomycin (ST488-2; Beyotime), 8-Br-cAMP (B5386; Sigma), MPA (B1510; APExBIO), 3-MA (3-methyladenine) (S24823; Yuanye Bio-Technology), TritonX-100 (T8200; Solarbio), Vimentin Monoclonal Antibody (60330-1; Proteintech), pULK1 S556 antibody (Bs-3464R; Bioss), p62/SQSTM Polyclonal Antibody (18420-1-AP; Proteintech), LC3 Polyclonal Antibody (14600-1-AP; Proteintech), AMPKα2 Rabbit pAb (A7339; ABclonal), pmTOR S2448 Rabbit pAb (AP0094; ABclonal), mTOR Rabbit pAb (A2445; ABclonal), ULK1 Rabbit pAb (A8529; ABclonal), Anti-AMPK alpha 1 (phospho T183) + AMPK alpha 2 (phospho T172) (ab23875; Abcam), pULK1 S757 Rabbit pAb (AP0736; ABclonal), GAPDH (A19056; ABclonal), Goat Anti-Mouse IgG Antibody (Ap124p; Millipore), Goat Anti-Rabbit IgG Antibody (Ap132p; Millipore), Alexa FluorTM 594 Goat Anti-Rabbit IgG (H + L) Antibody (A11037; Thermo), Alexa FluorTM 488 Goat Anti-Mouse IgG (H + L) Antibody (A11029; Thermo), TB Green Premix Ex Taq II (RR820A; TAKARA), PrimeScript RT reagent Kit with gDNA Eraser (RR047A; TAKARA).
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5

Quantitative Immunoblot Analysis

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Samples were boiled, and individual proteins were resolved by SDS-PAGE. After transfer, the PVDF membranes were decorated with the following primary antibodies: anti-HA antibody (BioLegend 901501, mouse), anti-TAP antibody (Invitrogen CAB1001, rabbit), anti-tubulin antibody (Abcam ab184970, rabbit), and anti-Sfh4 antibody (Bankaitis laboratory, Texas A&M University, rabbit). The blots were subsequently developed using the following HRP-conjugated secondary antibodies: goat anti-mouse IgG antibody (Millipore AP124P) and goat anti-rabbit IgG antibody (Millipore AP307P). Immunoblot images were acquired using Bio-Rad ChemiDoc XRS+ system. Relative band intensities on the blots were quantified using ImageJ (version 1.51s, National Institutes of Health).
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6

Steroid Hormone Measurement Protocol

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The P4 concentrations in the plasma were determined using the enzyme immuno-assay (EIA) methods described previously [19 (link)]. The
E2 levels in plasma were also measured using the EIA methods as described by Isobe [18 (link)], with some modifications. Briefly, 75
µl of extracted estradiol-17β standard or the sample and 25 µl of the 1:10,000 diluted estradiol-17β antibody (ab215528, Abcam, Cambridge, UK) were
applied to the wells of plates that had been previously coated with goat anti-rabbit IgG antibody (Millipore; Merck & Co., Inc., Kenilworth, NJ, USA). After 1 hr incubation, 25
µl of 1:2,500 diluted estradiol-6-CMO-HRP antigen (East Coast Bio, Inc., North Berwick, ME, USA) was added to the wells. The plate was incubated for another hour and then
washed six times. The substrate solution was applied to the wells, and then, they were incubated for 30 min at room temperature. The reaction was stopped by the addition of 1 M
H3PO4. The optical density was calculated at 450 nm. Sensitivity was 1.6 pg/ml for E2 and 0.5
ng/ml for P4. The intra-assay and inter-assay CVs were 11.4% and 6.7% for E2 and 8.6% and 3.2% for P4, respectively.
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7

Western Blot and qRT-PCR Reagents

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TRI Reagent, DNase I, and bovine serum albumin (BSA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Rabbit monoclonal anti-Tsc2 antibody was purchased from Novus Biologicals (Centennial, CO, USA). Mouse monoclonal anti-VAMP1/2 antibody, rabbit polyclonal anti-phospho-synapsin (Ser62/67) antibody, and mouse monoclonal anti-syntaxin-1 were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Rabbit polyclonal anti-synaptotagmin-1 antibody and rabbit monoclonal anti-SNAP25 antibody were obtained from Cell Signalling (Beverly, MA, USA). Rabbit polyclonal anti-GAPDH antibody and goat anti-rabbit IgG antibody were purchased from Sigma-Aldrich (St. Louis, MO, USA). Sheep anti-mouse IgG antibody was purchased from GE Healthcare (Little Chalfont, Buckinghamshire, UK). The chemiluminescent reagent Clarity Western ECL Substrate was purchased from Bio-Rad Laboratories (Hercules, CA, USA). The cOmplete protease inhibitor cocktail was purchased from Roche Diagnostics GmbH (Mannheim, Germany). Reagents for reverse transcription (High Capacity cDNA Reverse Transcription Kit) and PCR (TaqMan Fast Advanced Master Mix) were obtained from Thermo Fisher Scientific (Paisley, UK). DMSO and all the other common reagents were from Sigma-Aldrich (St. Louis, MO, USA).
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8

Antibody Characterization for Cell Analysis

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Antibodies used in this study were listed as follows: anti-p53 monoclonal antibody (1C12, Cell Signaling Technology), anti-p53 monoclonal antibody (7F5, Cell Signaling Technology), anti-Flag monoclonal antibody (M2, Sigma), anti-NS1 monoclonal antibody (NS1-23-1, Santa Cruz), anti-MDM2 polyclonal antibody (C18, Santa Cruz), anti-RPL11 polyclonal antibody (SAB2700779, Sigma), anti-RPL23 polyclonal antibody (I-20, Santa Cruz), anti-nucleolin monoclonal antibody (C23, Santa Cruz), anti-nucleolin polyclonal antibody (N2662, Sigma), anti-RPA194 monoclonal antibody (C-1, Santa Cruz), anti-β-actin monoclonal antibody (AC-15, Sigma), a horseradish peroxidase (HRP) conjugated goat anti mouse IgG antibody (Sigma), goat anti-rabbit IgG antibody (Sigma).
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9

Autophagy-related Proteins Modulation

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Mycophenolic acid (MPA), tunicamycin, and bafilomycin A1 were purchased from Sigma-Aldrich China (Shanghai, China). The primary antibodies, including rabbit anti-β-actin, mouse anti-LC3 antibody, rabbit anti-p62 antibody, mouse anti-ATG3 antibody, mouse anti-ATG5 antibody, mouse anti-ATG 7 antibody and the secondary antibodies, including goat anti-mouse IgG antibody, goat anti-rabbit IgG antibody were obtained from Sigma-Aldrich China (Shanghai, China). Mouse anti-HCV core antibody was purchased from Thermo Fisher Scientific China (Shanghai, China). Plasmids pCMV-myc-Atg3, pCMV-myc-Atg5, pCMV-myc-Atg7 were obtained from Addgene (www.addgene.org, MA, USA).
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10

Western Blot Analysis of Protein Targets

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Tissues were lysed in 2× Laemmli buffer (head, thorax and fat body) and proteins denatured at 95 °C for 5 min. Proteins from gut were extracted using 20% trichloric acid, washed in 1 M Tris buffer (no pH change), resuspended in 2× Laemmli buffer and denatured at 95 °C for 5 min. Proteins (10 μg) were separated using pre-stained SDS–PAGE gels (Bio-Rad) and wet-transferred onto a 0.45-μm nitrocellulose membrane (GE Healthcare). Blots were incubated with primary p-T389-S6K (CST, 9209), S6K2, Atg8 and Ref-2-P51 (link) antibodies (all diluted in a 1:1,000 ratio). Horseradish peroxidase-linked secondary antibodies, Goat Anti-Rabbit IgG Antibody (Sigma, 12-348; 1:10,000 dilution) or Goat Anti-Mouse IgG Antibody (Sigma, 12-349; 1:10,000 dilution) were used. Signal was developed using ECL Select Western Blotting Detection Reagent (GE Healthcare). Images were captured using a ChemiDoc XRS+ System with Image Lab (v5.1, Bio-Rad) and band intensity was analyzed using Fiji (v2.1.0).
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