Antibodies used for IB are as follows: t-IRS1 (catalog 2390), Insulin Receptor β (IRβ) (catalog 3025), IGF1R (catalog 3027), p-CaMKII (catalog 3356), t-CaMKII (catalog 4436), p-Erk (catalog 9101), t-Erk (catalog 4695), p-Akt (catalog 4060), t-Akt (catalog 9272), and NT (catalog 9691) were purchased from Cell Signaling Technology. Anti-β-actin (sc-1616), goat anti-mouse (sc-2031), and anti-rabbit IgG (sc-2004) were purchased from Santa Cruz Biotechnology. Anti-p-Tyr in IRS1 (Tyr608) (catalog 09-432) was obtained from Sigma-Aldrich and antibodies for IRS1 (catalog 09-248) and Galectin-3 (MABT51) were purchased from Millipore. Anti-VCAM1 (AF643) was obtained from R&D Systems. Antibodies for p-eNOS (catalog 612392) and t-eNOS (catalog 610297) were purchased from BD Biosciences. Endothelin Receptor type B Ab (NBP1-30599) was obtained from Novus Biologicals. Anti-α-actin (catalog A2547), L-NAME (N5751), EDN1 (E7764), wortmannin (W3144), PD98059 (P215), and BQ-788 (B157) were purchased from Sigma-Aldrich. Endothelin-1 ELISA kit (ADI-900-020A) was purchased from Enzo life sciences, and cGMP Direct Biotrak EIA (catalog 45-001-771) was purchased from GE Healthcare Life Sciences. DAF-2DA (catalog 251505) was purchased from EMD Millipore Chemicals. Fluo-4 NW Calcium Assay Kit (F36206) was purchased from (Invitrogen). Human Ox-LDL (BT-910) was purchased from Alfa Aesar.
Anti rabbit igg sc 2004
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Anti-rabbit IgG (sc-2004) is a secondary antibody designed for detection and labeling of rabbit primary antibodies in various immunodetection applications. It is a purified immunoglobulin fraction from goat serum.
Automatically generated - may contain errors
Lab products found in correlation
Anti mouse igg sc 2005, by Santa Cruz Biotechnology (2 mentions)
P erk, by Cell Signaling Technology (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
Insulin receptor β, by Cell Signaling Technology (1 mentions)
Anti α actin, by Merck Group (1 mentions)
P camkii, by Cell Signaling Technology (1 mentions)
Endothelin 1 elisa kit, by Enzo Life Sciences (1 mentions)
T enos, by BD (1 mentions)
Human ox ldl bt 910, by Thermo Fisher Scientific (1 mentions)
Cgmp direct biotrak eia, by GE Healthcare (1 mentions)
T camkii, by Cell Signaling Technology (1 mentions)
Igf 1r, by Cell Signaling Technology (1 mentions)
T akt, by Cell Signaling Technology (1 mentions)
T erk, by Cell Signaling Technology (1 mentions)
Protease inhibitor, by Roche (1 mentions)
Phosphatase inhibitor, by Merck Group (1 mentions)
Gtx113309, by GeneTex (1 mentions)
Sab2101014, by Merck Group (1 mentions)
Gt114, by GeneTex (1 mentions)
Gtx127375, by GeneTex (1 mentions)
7 protocols using anti rabbit igg sc 2004
1
Immunoblotting Assay for Endothelial Signaling
2
Protein Extraction and Western Blot Analysis
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For protein extraction, cells were washed twice with ice-cold PBS and lysed on ice in Golden lysis buffer (20 mM Tris-HCl (pH 8.0), 137 mM NaCl, 5.95 mM EDTA, 5 mM EGTA, 10 mM NaF, 1% Triton X-100, and 10% glycerol) supplemented with protease inhibitors (Roche, Indianapolis, IN, USA) and phosphatase inhibitors (Sigma-Aldrich, St. Louis, MO, USA). The proteins were separated via 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to polyvinylidene fluoride membranes. Specific antibodies against HAS3 (SAB2101014, Sigma-Aldrich, St. Louis, MO, USA), p21 (GT8611, GeneTex, Irvine, CA, USA), α-tubulin (GT114, GeneTex, Irvine, CA, USA), ATG5 (GTX113309, GeneTex, City, CA, USA), LC3 (GTX127375, GeneTex, Irvine, CA, USA), and GAPDH (sc-47724, Santa Cruz Biotechnology, Dallas, TX, USA) were diluted 1:2000 in Tris-buffered saline/Tween 20, and the membranes were incubated for 2 h at room temperature. Horseradish peroxidase-conjugated anti-mouse IgG (sc-2354, Santa Cruz Biotechnology, Dallas, TX, USA) and anti-rabbit IgG (sc-2004, Santa Cruz Biotechnology, Dallas, TX, USA) secondary antibodies were diluted 1:4000 and incubated with the membranes for 1 h at room temperature.
3
Western Blot Analysis of Myoclonin1
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Brain and lung (N = 1 WT and 1 Efhc1−/−) samples, cultured NSC (N = 1 WT and 2 Efhc1−/−) and HEK cells were homogenized in ice-cold 1X phosphate-buffered saline (PBS) supplemented with protease inhibitors (Complete, Roche). The following primary antibodies were used: mouse monoclonal anti-myoclonin1 (6A3-mAb, 1:2000 dilution) or rabbit polyclonal anti-myoclonin1 (mRib72-pAb, 1:200 dilution; kind gift from Prof. Ritsu Kamiya, University of Tokyo, Japan). HRP-conjugated anti-mouse IgG (W402B, Promega, 1:5000 dilution) or anti-rabbit IgG (sc-2004, SANTA CRUZ, 1:2000 dilution) were used for secondary antibody. Labeled proteins were revealed by using enhanced chemiluminescence (ECL) detection (Perkin-Elmer). Membranes were then washed with Restore Plus Western Blot stripping buffer (Thermo Scientific), re-probed with rabbit polyclonal anti-GAPDH (sc-25778, SANTA CRUZ, 1:1000 dilution) and HRP-conjugated anti-rabbit IgG antibody and revealed as described above.
4
Antibody Characterization and Validation
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The following primary antibodies were used: rat anti-HA (clone 3F10, Roche), rabbit anti-MyD88 (#3699S), rabbit anti-p105 (#4717), rabbit anti-IκBα (44D4) (#4812S), rabbit anti-p-IκBα (Ser32) (#2859) (all from Cell Signaling), mouse anti-V5 (Thermo Scientific, #R96025), rabbit anti-BANK1 (HPA037002), mouse anti β-actin (clone AC-15) (A5441) (both from Sigma-Aldrich), mouse anti-BANK1 (F-8) (sc-393611), rabbit anti-TRAF6 (H-274) (sc-7221), mouse anti-TRAF6 (D-10) (sc-8409), goat anti-TLR7 (V-20) (sc-16245), goat anti-TLR9 (N-15) (sc-13215), and mouse anti-Myc (9E10) (sc-40) antibodies; as a nonspecific control, the immunoglobulins rabbit IgG (sc-2027) and mouse IgG (sc-2025) (all from Santa Cruz) were used. The HRP-linked secondary antibodies for western blotting were anti-rabbit IgG (sc-2004) (#7074), anti-mouse IgG (sc-2005) (#7076) (both from Santa Cruz and Cell Signaling), anti-goat IgG (sc-2354) or anti-rat IgG (sc-2032) (both from Santa Cruz). The secondary antibodies for IF reactions were Alexa Fluor 555 donkey anti-goat IgG (H + L) (#A-21432), Alexa Fluor 647 goat anti-mouse/rabbit IgG (H + L) (#A-21235; #A-31634) and Alexa Fluor 488 goat anti-rabbit/mouse IgG (H + L) (#A-11034; #A-11001) (all from Thermo Scientific).
5
Antibody Detection for EMT Markers
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Primary antibodies against AGO2 (ab186733) and Survivin (ab469) were purchased from Abcam Inc. (Cambridge, UK). Antibodies for detecting Snail (#3895) and Vimentin (#5741) were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Primary antibody for GAPDH (sc-25778) and secondary antibodies including anti-rabbit IgG (sc-2004) and anti-mouse IgG (sc-2005) were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA).
6
Antibody Sources for Protein Analysis
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The anti-p65 (sc-109), anti-IκBα (sc-371), anti-RANKL (sc-7628) and anti-rabbit IgG (sc-2004) antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The anti-phosphorylated p65 (Serine 536) (#3031) and anti-mouse IgG antibodies were obtained from Cell Signaling (Beverly, MA, USA) and Amersham Bioscience (Piscataway, NJ, USA), respectively. The anti-rat matrix metalloproteinase-9 (MMP-9) antibody (AB19016) was obtained from Millipore (Billerica, MA, USA). The anti-β-actin antibody (AC-15) was purchased from Sigma-Aldrich (St. Louis, MO, USA).
7
AMPKα Activation and PDH Regulation
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All primary and secondary antibodies were purchased from commercial sources, listed as follows: AMPKα Antibody (2532, Cell Signaling, Danvers, MA, USA), phospho-AMPKα (Thr172) (2535, Cell Signaling), Anti-Pyruvate Dehydrogenase E1-alpha subunit antibody (ab110334, Abcam, Cambridge, UK), Anti-PDHA1 (phospho S293) antibody (ab177461, Abcam), β-Actin (A5441, Sigma-Aldrich), α-Tubulin (sc-8035, Santa Cruz Biotechnology, Santa Cruz, CA, USA). The secondary antibodies for horseradish peroxidase (HRP) detection were anti-rabbit IgG (sc-2004, Santa Cruz Biotechnology) and anti-mouse IgG (PI-2000, Vector Laboratories, Burlingame, CA, USA).
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