Hrp conjugated anti rabbit igg 7074

Manufactured by Cell Signaling Technology

Sourced in United States

HRP-conjugated anti-rabbit IgG (7074S) is a laboratory reagent used for detection and quantification of rabbit immunoglobulin G (IgG) in various immunoassays. It is conjugated with horseradish peroxidase (HRP), an enzyme that can catalyze a color-producing reaction, enabling visualization and measurement of the target protein.

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Lab products found in correlation

Mouse anti gapdh nb300 221 antibody, by Novus Biologicals (2 mentions) Ab227036, by Abcam (2 mentions) Ab175430, by Abcam (2 mentions) Hrp conjugated anti mouse igg 1706516 antibody, by Bio-Rad (2 mentions) Mg132, by Targetmol (1 mentions) Dual luciferase reporter gene assay kit, by Promega (1 mentions) Pericyte medium pm, by ScienCell (1 mentions) Alexa fluor 594 conjugated donkey anti rabbit igg ab150064, by Abcam (1 mentions) Cycloheximide (chx), by Targetmol (1 mentions) Chip pcr kit, by Merck Group (1 mentions) Pkh26, by Merck Group (1 mentions) Pkh67, by Merck Group (1 mentions) Matrigel, by BD (1 mentions) Endothelial cell medium (ecm), by ScienCell (1 mentions) Recombinant human tnfα 210 ta, by R&D Systems (1 mentions) U0126 9903, by Cell Signaling Technology (1 mentions) Sb203580 5633, by Cell Signaling Technology (1 mentions) Ab9829, by Abcam (1 mentions) Ab31630, by Abcam (1 mentions) Ab64693, by Abcam (1 mentions)

7 protocols using hrp conjugated anti rabbit igg 7074

Angiogenesis Regulation in Endothelial Cells

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Endothelial cell medium (ECM) and pericyte medium (PM) were products of ScienCell Research Laboratories (San Diego, CA, USA). Antibodies against Ki67 (#12,202), α-SMA (#19,245), bFGF (#20,102), FGFR1 (#76,123), FGFR2 (#23,328), β-actin (#23,328) and HRP-conjugated anti-rabbit IgG (#7074) were obtained from Cell Signaling Technology (Danvers, MA, USA). Antibodies against Gli1 (AF3455) and CD31 (AF3628), and a Proteome Profiler Human Angiogenesis Array Kit (ARY007) were obtained from R&D Systems (Minneapolis, MN, USA). Alexa Fluor 594-conjugated donkey anti-rabbit IgG (ab150064) and Alexa Fluor488- conjugated donkey anti-mouse IgG (ab150073) were obtained from Abcam (Cambridge, UK). GANT-61, cycloheximide (CHX) and MG132 were purchased from TargetMol (Shanhai, China). Matrigel was obtained from BD Biosciences (Franklin Lakes, NJ). Recombinant human bFGF was purchased from BioVision (Palo Alto, CA, USA). A ChIP PCR kit was obtained from Merck Millipore (Billerica, MA, USA). A dual luciferase reporter gene assay kit was provided by Promega (Madison, WI, USA). Rhodamine-labeled lysinated dextran (70-kDa) was product from Thermo Fisher Scientific (Waltham, MA, USA). PKH 26, PKH 67 and other reagents were obtained from Sigma-Aldrich (St. Louis, MO, USA).

Lei X., Li Z., Huang M., Huang L., Huang Y., Lv S., Zhang W., Chen Z., Ke Y., Li S., Chen J., Yang X., Deng Q., Liu J, & Yu X. (2024). Gli1-mediated tumor cell-derived bFGF promotes tumor angiogenesis and pericyte coverage in non-small cell lung cancer. Journal of Experimental & Clinical Cancer Research : CR, 43, 83.

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Signaling Pathways Activation in TNF-α and CCL21 Response

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Recombinant human TNF‐α (210‐TA) and CCL21 (366‐6C) were purchased from R&D Systems (Minneapolis, MN, USA). The following antibodies were used in Western blot analyses and immunocytochemistry: rabbit anti‐TNF‐α (ab6671), goat anti‐rabbit FITC‐conjugated IgG H&L (ab6717), and rabbit anti‐CCR7 (ab103404) from Abcam (Cambridge, MA, USA); rabbit anti‐p38 (8690), rabbit anti‐phospho‐p38 (Thr180/Tyr182; 4511), rabbit anti‐ERK (9102), rabbit anti‐phospho‐ERK (Thr202/Tyr204; 9101), and HRP‐conjugated anti‐rabbit IgG (7074) antibodies from Cell Signaling Technology (Danvers, MA, USA); and mouse anti‐GAPDH (NB300‐221) antibody from Novus Biologicals (Littleton, CO, USA). The ERK inhibitor U0126 (9903) and the p38 inhibitor SB203580 (5633) were purchased from Cell Signaling Technology.

Maolake A., Izumi K., Natsagdorj A., Iwamoto H., Kadomoto S., Makino T., Naito R., Shigehara K., Kadono Y., Hiratsuka K., Wufuer G., Nastiuk K.L, & Mizokami A. (2018). Tumor necrosis factor‐α induces prostate cancer cell migration in lymphatic metastasis through CCR7 upregulation. Cancer Science, 109(5), 1524-1531.

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Characterization of CCL20-Mediated Signaling

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Recombinant human CCL20 (360-MP) was purchased from R&D Systems (Minneapolis, MN, USA). The following antibodies were used in western blot analyses, neutralizing assays, and immunohistochemistry: rabbit anti-CCL20 (ab9829), rabbit anti-CCR6 (ab227036), rabbit anti-CCR7 (ab103404), rabbit anti-CD206 (ab64693), mouse anti-CD68 (ab31630), mouse anti-Snail (ab117866), mouse anti-Twist (ab175430), and rabbit anti-Vimentin (ab92547), from Abcam (Cambridge, MA, USA); rabbit anti-Akt (9272S), rabbit anti-phospho-Akt (Ser473; 9271S), and HRP-conjugated anti-rabbit IgG (7074) antibodies from Cell Signaling Technology (Danvers, MA, USA); mouse anti-GAPDH (NB300-221) antibody from Novus Biologicals (Littleton, CO, USA); and HRP-conjugated anti-mouse IgG (1706516) antibody from Bio-Rad Laboratories (Hercules, CA, USA). The Akt inhibitor AZD5363 (S8019) was purchased from Selleck Chemicals (Houston, TX, USA).

Kadomoto S., Izumi K., Hiratsuka K., Nakano T., Naito R., Makino T., Iwamoto H., Yaegashi H., Shigehara K., Kadono Y., Nakata H., Saito Y., Nakagawa-Goto K, & Mizokami A. (2019). Tumor-Associated Macrophages Induce Migration of Renal Cell Carcinoma Cells via Activation of the CCL20-CCR6 Axis. Cancers, 12(1), 89.

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Western Blot Analysis of Apoptosis Regulators

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Cells were harvested in a cell lysis buffer (50 mM Tris-HCl, 150 mM NaCl, 1% Triton X-100, 0.1% SDS, and 1 mM PhCH₂SO₂F) and a cocktail of protease inhibitors (Roche, IN). Total protein (50 μg) was electrophoresed on a 12% SDS-polyacrylamide gel and transferred to polyvinylidene difluoride membranes (Millipore, MA). Membranes were blocked at room temperature for 1 h with a blocking buffer (5% non-fat dry milk in 10 mM Tris-HCl pH 7.6, 150 mM NaCl, 0.1% Tween-20) and then incubated at 4°C overnight with rabbit anti-MCL1 (5453S), anti-BCL-2 (2870S), anti-β-actin (4970S) (Cell signaling technology, MA), anti-BCL-X_L/BCL-X_S (sc-634) (Santa Cruz, CA), or mouse anti-SF3B1 (D221-3) (MBL, IL) followed by 1-h incubation with a horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (7074S) or anti-mouse secondary antibodies (Cell signaling technology, MA). Blots were developed with ECL Plus reagents (PerkinElmer Life and Analytical Science, MA) and quantified using an Image Gauge Version 4.0 (FUJIFILM, NJ).

Gao Y., Trivedi S., Ferris R.L, & Koide K. (2014). Regulation of HPV16 E6 and MCL1 by SF3B1 inhibitor in head and neck cancer cells. Scientific Reports, 4, 6098.

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Antibody Source and Dilution Protocol

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Antibodies against the following proteins/epitopes were purchased from the indicated sources: GAPDH (sc-47724), His tag (sc-8036), GST (sc-138), p53 (DO1, sc-126), MDM2 (sc-965), DAXX (sc-8043), α-synuclein (syn211, sc-12767) (Santa Cruz Biotechnology); Flag tag (14793) and DAXX (4533S) (Cell Signaling Technology); p53 (PAb1620, OP33; PAb240, OP29), MDM2 (OP46) (Calbiochem); HA tag (ab137838) and luciferase (ab21176) (Abcam); and β-Amyloid 1–42 (Aβ42, 805509) (BioLegend). HRP-conjugated anti-rabbit IgG (7074S) and anti-mouse IgG (7076S) antibodies were purchased from Cell Signaling Technology; IRDye 800CW (926–32211, anti-Rabbit) and IRDye 680RD (926–68070, anti-Mouse) secondary antibodies from Li-Cor; anti-Flag M2 Affinity Gel (A2220), 3xFlag peptide (F4799), firefly luciferase (L9420), and Aβ42 (A9810) from Sigma Aldrich; HSP70 (human HSP72, ADI-NSP-555), HSP40 (human HDJ1, ADI-SPP-400), and ATP regeneration solution (BML-EW9810–0100) from Enzo Life Sciences; and 6xHis-ubiquitin (U-530), UBE1 (E-304), and UBE2D2 (E2–622) from Boston Biochem. α-Synuclein (RP-003, RP-001) was purchased from Proteos. For western blot, anti-DAXX, p53, MDM2, and α-synuclein antibodies were used at 1:1,000 dilution, IRDye 800CW and IRDye 680RD at 1:10,000 dilution, and all other antibodies at 1:2,000 dilution.

Huang L., Agrawal T., Zhu G., Yu S., Tao L., Lin J., Marmorstein R., Shorter J, & Yang X. (2021). DAXX represents a new type of protein-folding enabler. Nature, 597(7874), 132-137.

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EMT Evaluation via Western Blot

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On day 3 after seeding on a polyacrylamide gel, we prepared cell lysates by scraping the gel surface and detected the level of proteins by western blot analysis. To determine the degree of the EMT, we compared the level of epithelial and mesenchymal marker proteins, including E-cadherin, vimentin, and N-cadherin. GAPDH was used as a loading control. Following primary and secondary antibodies were purchased from Cell Signaling Technology: E- cadherin (3195s), Vimentin (5741s), N-Cadherin (13116s), GAPDH (5174s) and HRP-conjugated anti-rabbit IgG (7074s). We used Image J to quantify the level of target proteins and to compare their levels between cell lines.

Kim J.H., Pegoraro A.F., Das A., Koehler S., Ujwary S.A., Lan B., Mitchel J.A., Atia L., He S., Wang K., Bi D., Zaman M., Park J.A., Butler J.P., Lee K.H., Starr J.R, & Fredberg J.J. (2019). Unjamming and collective migration in MCF10A breast cancer cell lines. Biochemical and biophysical research communications, 521(3), 706-715.

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Investigating Cellular Signaling Pathways

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Kahweol acetate (sc-228383A) and cafestol (sc-204663) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). The following antibodies were used during western blot analyses: mouse anti-Bcl-2 (15071S), rabbit anti-Bcl-xL (2764S), rabbit anti-STAT3 (8719S), rabbit anti-phospho-STAT3 (9131S), rabbit anti-Akt (9272S), rabbit anti-phospho-Akt (Ser473; 9271S), rabbit anti-ERK (9102S), rabbit anti-phospho-ERK (Thr202/Tyr204; 9101S), rabbit anti-PD-L1 (13684S), and horseradish peroxidase (HRP)-conjugated anti-rabbit IgG (7074S) antibodies from Cell Signaling Technology (Danvers, MA, USA); mouse anti-Snail (ab117866), mouse anti-Twist (ab175430), rabbit anti-CCR2 (ab155321), rabbit anti-CCR5 (ab32048), and rabbit anti-CCR6 (ab227036) from Abcam (Cambridge, MA, USA); mouse anti-GAPDH (NB300-221) from Novus Biologicals (Littleton, CO, USA); and HRP-conjugated anti-mouse IgG (1706516) antibody from Bio-Rad Laboratories (Hercules, CA, USA). Antibody information used in western blot analyses for Supplementary Figures is shown in Supplementary Figure S6.

Makino T., Izumi K., Hiratsuka K., Kano H., Shimada T., Nakano T., Kadomoto S., Naito R., Iwamoto H., Yaegashi H., Shigehara K., Kadono Y., Nakata H., Saito Y., Nakagawa-Goto K., Sakai N., Iwata Y., Wada T, & Mizokami A. (2021). Anti-proliferative and anti-migratory properties of coffee diterpenes kahweol acetate and cafestol in human renal cancer cells. Scientific Reports, 11, 675.

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