Peroxidase conjugated affinipure goat anti mouse igg h l

Manufactured by Jackson ImmunoResearch

Sourced in United States

Peroxidase-conjugated AffiniPure goat anti-mouse IgG (H + L) is a secondary antibody conjugated with the enzyme peroxidase. It is designed to detect and bind to mouse immunoglobulin G (IgG) antibodies, including both the heavy and light chains.

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Lab products found in correlation

Peroxidase conjugated affinipure goat anti rabbit igg h l, by Jackson ImmunoResearch (5 mentions) Hif 1α, by Cell Signaling Technology (2 mentions) Mini protean tgx precast protein gel, by Bio-Rad (1 mentions) Mjfr 19 7 8, by Abcam (1 mentions) Can get signal, by Toyobo (1 mentions) Protran, by Cytiva (1 mentions) Phosphatase inhibitor, by Merck Group (1 mentions) Gapdh, by Merck Group (1 mentions) Sds page loading buffer, by Beyotime (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Sc 390811, by Santa Cruz Biotechnology (1 mentions) Immobion p transfer membrane, by Merck Group (1 mentions) Gapdh antibody, by Boster Bio (1 mentions) Caspase 3 19677 1 ap, by Proteintech (1 mentions) Fluorescein fitc conjugated affinipure goat anti rabbit igg h l, by Jackson ImmunoResearch (1 mentions) Cy3 conjugated affinipure goat anti rabbit igg h l, by Jackson ImmunoResearch (1 mentions) Pvdf membrane, by Merck Group (1 mentions) Radioimmunoprecipitation assay (ripa), by Cell Signaling Technology (1 mentions) β actin, by Abcam (1 mentions) Runx2, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

Peroxidase AffiniPure Goat Anti-Mouse IgG (H+L) (42 citations) Peroxidase AffiniPure Goat Anti-Mouse IgG (42 citations) AffiniPure goat anti-mouse IgG (H+L) (38 citations) Goat anti-mouse IgG (H+L) (36 citations) AffiniPure goat anti-mouse IgG (28 citations) Peroxidase-conjugated AffiniPure goat anti-mouse IgG (16 citations) Peroxidase-conjugated goat anti-mouse IgG (H+L) (11 citations) Peroxidase-conjugated AffiniPure goat anti-mouse (4 citations) Normal IgG/Peroxidase-conjugated AffiniPure Goat Anti-Rabbit/Mouse IgG (H+L) (4 citations) Peroxidase-conjugated AffiniPure F(ab’)2 fragment goat anti mouse IgG (H+L) (2 citations)

11 protocols using peroxidase conjugated affinipure goat anti mouse igg h l

Western Blot Analysis of LRRK2 Phosphorylation

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Samples were loaded on 4–20% Mini-PROTEAN® TGX™ Precast Protein Gels, 15-well, 15 µl (Bio-rad, #4561096), then proteins were transferred onto Amersham™ Protran™ NC Nitrocellulose Membranes (GE Healthcare, 45-004-001). Membranes were blocked by 1% bovine serum albumin in Tris-buffered saline with 0.1% Tween 20 (TBST). The immunoreaction enhancer solution Can Get Signal™ (TOYOBO, NKB-101) was used for the application of the primary and secondary antibodies listed in Table 3.

Table 3.

Summary of used antibodies for western blot.

Antibody name	Dilution
Recombinant Anti-LRRK2 (phospho S1292) rabbit monoclonal antibody [MJFR-19-7-8] (abcam, sb203181)	1:500
Anti-c-Myc mouse monoclonal antibody [9E10] (Santa Cruz, sc-40)	1:2000
Anti-LRRK2/Dardarin mouse monoclonal antibody [N241A/34] (Neuromab, 75-253)	1:1000
Anti-β-actin mouse monoclonal antibody [C4] (Santa Cruz. Sc-47778)	1:4000
Anti-α-tubulin mouse monoclonal antibody [B-5-1-2] (Sigma-aldrich, T5168)	1:10000
Peroxidase-conjugated AffiniPure Goat Anti-Mouse IgG (H + L) (#115-035-003; Jackson Immunoresearch Laboratories Inc.)	1:5000
Peroxidase-conjugated AffiniPure Goat Anti-Rabbit IgG (H + L) (#111-035-144; Jackson Immunoresearch Laboratories Inc.).	1:5000

Nam D., Kim A., Han S.J., Lee S.I., Park S.H., Seol W., Son I, & Ho D.H. (2021). Analysis of α-synuclein levels related to LRRK2 kinase activity: from substantia nigra to urine of patients with Parkinson’s disease. Animal Cells and Systems, 25(1), 28-36.

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Quantifying Gd-IgA1-specific IgA Antibodies

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ELISA plates were coated with Fab fragment of Gd-IgA1 (Ste) described above [15] (link). Captured antibodies were detected by incubation with mouse monoclonal antibody to human IgA (Fc-specific) (Applied Biological Materials Inc., Richmond, BC) and detected by Peroxidase-conjugated AffiniPure Goat Anti-Mouse IgG (H+L) (Jackson Immuno Research, West Grove, PA). Serum levels of Gd-IgA1-specific IgA were expressed in Units (1 unit as OD 1.0 measured at 490 nm).

Yanagawa H., Suzuki H., Suzuki Y., Kiryluk K., Gharavi A.G., Matsuoka K., Makita Y., Julian B.A., Novak J, & Tomino Y. (2014). A Panel of Serum Biomarkers Differentiates IgA Nephropathy from Other Renal Diseases. PLoS ONE, 9(5), e98081.

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Immunoblotting protocol for GCLC protein analysis

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For immunoblotting, cells were washed with cold PBS for three times and lysed in the RIPA extraction regent (Pierce Biotechnology, Rockford, IL, USA) containing 1% protease inhibitor cocktail (Roche, France) and phosphatase inhibitor (Sigma, France) for 1 hour on ice. After clarified by centrifugation for 10min at 12,000×g, the supernatant of cell lysates was added with 5X SDS-PAGE loading buffer (Beyotime, Shanghai, China. Cat No#P0015) and then heated to 95-100°C for 10 minutes. 20µg total protein were loaded per well for blotting and electrotransfer to 0.45µm PVDF membrane (Immobion-P Transfer Membrane, Millipore, Billerica, MA, USA. Cat No# IPVH 00010). Following 5% non-fat dry milk blockage of membranes, specific primary antibody for GCLC (1:1000, sc-390811, Santa Cruz, CA, USA) and GAPDH (1:2000; Sigma-Aldrich, USA. Cat No# G9295) serving as internal reference were incubated overnight in 4°C. After washed three times for 5 minutes with TBST, the membranes were incubated with secondary antibody (Peroxidase-conjugated AffiniPure Goat anti-Mouse IgG (H+L), 1:2000; Jackson ImmunoResearch, USA. Cat No#111-035-003) for 2 hours and then exposed to Tanon-5200 Chemiluminescent Imaging System (Tanon, China) for imaging. All the experiments were performed for three times.

Sun J., Zhou C., Ma Q., Chen W., Atyah M., Yin Y., Fu P., Liu S., Hu B., Ren N, & Zhou H. (2019). High GCLC level in tumor tissues is associated with poor prognosis of hepatocellular carcinoma after curative resection. Journal of Cancer, 10(15), 3333-3343.

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Antibodies and Reagents for Western Blot

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Primary antibodies against AEG-1/MTDH (13860-1-AP), E-cadherin (1702-1), N-cadherin (2447-1), and vimentin (2862-1) were obtained from Epitomics Inc. (Burlingame, CA, USA). Primary antibodies against poly-ADP-ribose polymerase (PARP) (BS70001), PI3K (BS3006), Bax (BS1030), Bcl-2 (BS1031), and C-myc (BS2462) were obtained from Bioworld Technology Inc. (Minneapolis, MN, USA). Primary antibodies against NF-κB-p65 (10745-1-AP), caspase-8 (13423-1-AP), caspase-9 (10380-1-AP), and caspase-3 (19677-1-AP) were obtained from Proteintech (Wuhan, China); Primary antibodies against Lamin A (sc-177452), AKT (sc-8312), p-AKT (sc-7985), ERK (sc-154), and p-ERK (sc-23759) were obtained from Santa Cruz Biotechnology (Santa Cruz Biotechnology Inc., CA). The GAPDH antibody was purchased from Boster (Wuhan, China). The secondary antibodies peroxidase-conjugated AffiniPure Goat Anti-Mouse IgG (H+L) (111-035-003), peroxidase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) (111-035-003), Cy™ 3-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (111-165-003), and Fluorescein (FITC)-conjugated AffiniPure Goat Anti-Rabbit IgG(H+L) (111-095-003) were obtained from Jackson Immuno Research Laboratories, Inc. (USA). Temozolomide (85622-93-1) was obtained from Meilun Biology Technology (Dalian, China).

Zou M., Duan Y., Wang P., Gao R., Chen X., Ou Y., Liang M., Wang Z., Yuan Y., Wang L, & Zhu H. (2016). DYT-40, a novel synthetic 2-styryl-5-nitroimidazole derivative, blocks malignant glioblastoma growth and invasion by inhibiting AEG-1 and NF-κB signaling pathways. Scientific Reports, 6, 27331.

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Osteogenic Differentiation of hADSCs

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After culturing hADSCs in different concentrations of LL37 for 7 days, RIPA (Cell Signaling Technology) and loading buffers (Takara) were used to lyse hADSCs. An SDS-PAGE gel (12%) was prepared and used for the electrophoretic separation of the proteins at 80 V for 30 min, and then 120 V for 30 min. After electrophoresis, the gel was transferred to a 0.22 μm polyvinylidene fluoride (PVDF) membrane (Millipore, Billerica, MA, United States) and blocked in 5% BSA for 1 h at room temperature. The PVDF membrane was incubated with primary antibodies against Col1, Opn, Ocn, Runx2, Bsp (Santa Cruz Biotechnology, Inc.), and β-actin (Abcam, Cambridge, MA, United States) overnight at 4°C. TBST buffer was used to wash the PVDF membrane three times for 10 min before the membranes were probed with peroxidase-conjugated affinipure goat anti-mouse IgG (H + L) (Jackson ImmunoResearch Laboratories, Inc., United States). Finally, the membrane was scanned using a Tanon-5200 image scanner (Tanon, China) and the relative gray levels of proteins were normalized by the β-actin gray level.

Li L., Peng Y., Yuan Q., Sun J., Zhuang A, & Bi X. (2021). Cathelicidin LL37 Promotes Osteogenic Differentiation in vitro and Bone Regeneration in vivo. Frontiers in Bioengineering and Biotechnology, 9, 638494.

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Protein Blot Analysis of Cellular Stress Markers

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For protein blot analyses, cells were grown to full confluence, washed twice with 1X PBS, immediately scraped off the plate into a microcentrifuge tube with 1X Laemmli sample buffer for CISD2 and mNT antibodies, and heated to 95°C for 10 minutes. For GPX4, TfR, TXNIP and TRX2, RIPA (Radioimmunoprecipitation assay; Sigma-Aldrich) buffer was used and lysates were kept on ice for 2 hours with vortex every 30 minutes, then centrifugation at 15,000 rpm for 15 minutes, supernatant was used for protein denaturation with Laemmli sample buffer 5X. Protein gels were loaded with equal amounts of protein, determined using The Pierce 660 nm Protein Assay (Thermo Scientific™, Cat. No. 22662) [11 (link)], and analyzed using antibodies against CISD2 and mitoNEET [23 , 33 (link)], TXNIP (Novus Biologicals, NBP2–27095), Thioredoxin 2 (Novus Biologicals, NBP1–92499), β-actin (R&D Systems, MAB8929), GPX4 (R&D Systems, Cat. No. MAB5457), and TfR (Abcam, ab84036). Goat Anti-Rabbit IgG, H & L Chain Specific Peroxidase Conjugate (Sigma, 401315) and Peroxidase-conjugated AffiniPure Goat anti-mouse IgG (H+L) (Jackson ImmunoResearch Laboratories, AB_10015289) were used as secondary antibodies (7, 24). All experiments were repeated at least 3 times.

Karmi O., Sohn Y.S., Zandalinas S.I., Rowland L., King S., Nechushtai R, & Mittler R. (2021). Disrupting CISD2 function in cancer cells primarily impacts mitochondrial labile iron levels and triggers TXNIP expression. Free radical biology & medicine, 176, 92-104.

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Antibodies Used in Hypoxia Signaling

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The following antibodies
were used in this study: ACTB (Merck; A5316), HIF1-α (Cell Signaling;
14179), HIF2/EPAS-1 (Santa Cruz Biotechnology sc-13596), HXKII (Santa
Cruz Biotechnology sc-6521), CAIX (Novus Biotechnology NB100-417),
SIRT3 (Cell Signaling; 2627S), MnSOD (Abcam ab13533), MnSOD K68 (Abcam
ab137037), MnSOD K122 (Abcam ab214675), peroxidase-conjugated AffiniPure
Goat Anti-Rabbit IgG (H + L) (Jackson ImmunoResearch; 111-035-003),
and peroxidase-conjugated AffiniPure Goat Anti-Mouse IgG (H + L) (Jackson
ImmunoResearch; 115-035-062).

Zwergel C., Aventaggiato M., Garbo S., Di Bello E., Fassari B., Noce B., Castiello C., Lambona C., Barreca F., Rotili D., Fioravanti R., Schmalz T., Weyand M., Niedermeier A., Tripodi M., Colotti G., Steegborn C., Battistelli C., Tafani M., Valente S, & Mai A. (2023). Novel 1,4-Dihydropyridines as Specific Binders and Activators of SIRT3 Impair Cell Viability and Clonogenicity and Downregulate Hypoxia-Induced Targets in Cancer Cells. Journal of Medicinal Chemistry, 66(14), 9622-9641.

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Neuronal Cell Culture Protocols

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ADMA was obtained from Sigma-Aldrich (St. Louis, MO, USA). Neurobasal A medium, B27 supplement, and no glucose DMEM were purchased from Invitrogen (Grand Island, NY, USA), and mouse anti-connexin 36 antibody was obtained from Invitrogen (Carlsbad, CA, USA). Mouse anti-β-tubulin III (neuronal) antibody was purchased from Sigma-Aldrich (St. Louis, MO, USA). Peroxidase-conjugated AffiniPure goat anti-mouse IgG (H+L) was purchased from Jackson ImmunoResearch (West Grove, PA, USA). A cytotoxicity detection kit based on lactate dehydrogenase (LDH) was obtained from Roche Diagnostic GmbH (Mannheim, Germany). 7-Nitroindazole (7-NI), 1400w, BCA Protein Assay Kit, and fluorescent probe 3-amino, 4-aminomethyl-2′,7′-difluorescein, diacetate (DAF-FM DA), and phenylmethanesulfonyl fluoride (PMSF) were purchased from Beyotime (Jiangsu, China). Other reagents were obtained from Sigma-Aldrich.

Fang K., Chen S., Wang Y., Chen F., Cui M, & Dong Q. (2022). Asymmetric Dimethylarginine Protects Neurons from Oxygen Glucose Deprivation Insult by Modulating Connexin-36 Expression. Oxidative Medicine and Cellular Longevity, 2022, 5339361.

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Antibody and Chemical Treatments for Autophagy Study

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The following antibodies were used in this study: acetylated lysine (Cell Signaling; Danvers, MA, USA; 9441), BECN1 (Cell Signaling, 3738), BNIP3 (Santa Cruz Biotechnology, Dallas, TX, USA; sc-56167), GAPDH (Santa Cruz Biotechnology; sc-137179), GAC (GeneTex, Irvine, CA, USA; GTX131263), HIF-1α (Cell Signaling; 14179), MAP1LC3B (Novus Biologicals, Centennial, CO, USA; NB600-1384), PRKN (Santa Cruz Biotechnology, sc-32282), SLC20A1/2 (PiT-1/2) (Santa Cruz Biotechnology, sc-101298), SLC25A3 (Santa Cruz Biotechnology, sc-376742), ULK1 (Abcam, Cambridge, UK; ab128859), VDAC1 (Santa Cruz Biotechnology, sc-390996), pULK1 (Abcam; ab156920), peroxidase-conjugated AffiniPure goat anti-rabbit IgG (H + L) (Jackson ImmunoResearch, West Grove, PA, USA; 111-035-003), and peroxidase-conjugated AffiniPure goat anti-mouse IgG (H + L) (Jackson ImmunoResearch; 115-035-062).
Lanthanum acetate (Merck; 306339) was dissolved in distilled water, filtered and then added to a final concentration of 2 mM every 24 h and 2 h before harvesting the cells.
Cobalt (III) chloride hexahydrate (Merck; C8661) was dissolved in distilled sterile water and added to a final concentration of 200 µM for 24 h or 48 h before harvesting the cells.
Bafilomycin A₁ (Merck, B1793) was dissolved in dimethyl sulfoxide (DMSO) and added to a final concentration of 100 nM for 2 h.

Barreca F., Aventaggiato M., Vitiello L., Sansone L., Russo M.A., Mai A., Valente S, & Tafani M. (2023). SIRT5 Activation and Inorganic Phosphate Binding Reduce Cancer Cell Vitality by Modulating Autophagy/Mitophagy and ROS. Antioxidants, 12(8), 1635.

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Western Blot Protein Analysis

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The targeting proteins were analyzed by standard Western blot methods and probed with following antibodies: anti-flag M2, monoclonal antibody (dilution = 1:5000, Sigma, F3165), Monoclonal anti-a-tubulin antibody (dilution = 1:5000, Sigma, T5168), Anti mCherry-tag mouse monoclonal antibody (Sungene Biotech, KM8017), Peroxidase-conjugated Affinipure Goat Anti-Rabbit IgG(H + L) (Jackson, 111-035-003), Peroxidase-conjugated Affinipure Goat Anti-Mouse IgG(H + L) (Jackson, 115-035-003).

He Y., Hao F., Fu H., Tian G., Zhang Y., Fu K, & Qi B. (2023). N-glycosylated intestinal protein BCF-1 shapes microbial colonization by binding bacteria via its fimbrial protein. Cell reports, 42(1).

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