Goat Anti-Mouse IgG H&L (HRP) preadsorbed

38 product images

• 

  Immunoprecipitation - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  Lane 1: Anti-GFP antibody (Magnetic beads) (Magnetic beads Anti-GFP VHH Single Domain antibody ab193983) + 10ul recombinant GFP (Recombinant A. victoria GFP protein ab84191)

  Lane 2: Anti-GFP antibody (Magnetic beads) (Magnetic beads Anti-GFP VHH Single Domain antibody ab193983) + 10ul GFP fusion protein.

  Lane 3: Off-target sdAb beads: Anti-Rabbit VHH Single Domain Antibody (Magnetic) + 10ul recombinant GFP (Recombinant A. victoria GFP protein ab84191)

  Lane 4: Anti-GFP antibody (Anti-GFP antibody ab290) + Protein A agarose + 10ul recombinant GFP (Recombinant A. victoria GFP protein ab84191)

  Lane 5: Anti-GFP antibody (Anti-GFP antibody ab290) + Protein A agarose + 10ul fusion GFP

  Anti-GFP antibody (Magnetic beads) (Magnetic beads Anti-GFP VHH Single Domain antibody ab193983) (20ul slurry) was used to pull down recombinant GFP (Recombinant A. victoria GFP protein ab84191) and GFP fusion protein. Anti-GFP antibody ab290 (Rabbit polyclonal to GFP - ChIP Grade) and Protein A agarose was used as a positive control. The gel was stained with Optiblot Blue (InstantBlue® Coomassie Protein Stain (ISB1L) ab119211).

  Lanes 1 - 2: Immunoprecipitation - Magnetic beads Anti-GFP VHH Single Domain antibody (Magnetic beads Anti-GFP VHH Single Domain antibody ab193983) at 20 µL

  Lane 3: Immunoprecipitation - Magnetic beads Anti-GFP VHH Single Domain antibody (Magnetic beads Anti-GFP VHH Single Domain antibody ab193983)

  Lanes 4 - 5: Immunoprecipitation - Magnetic beads Anti-GFP VHH Single Domain antibody (Magnetic beads Anti-GFP VHH Single Domain antibody ab193983) at 1/500 dilution

  Secondary

  All lanes: Immunoprecipitation - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/20000 dilution

  Observed band size: 27 kDa

  Exposure time: 30s

• 

  Immunoprecipitation - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  Lane 1: HEK293 cell lysate overexpressing GFP - 10ug

  Lane 2: Unbound IP supernatant: Anti-GFP antibody (Magnetic beads) + 100ug HEK293 cell lysate overexpressing GFP

  Lane 3: Negative IP: off-target sdAb beads: Anti-GFP antibody (Magnetic beads) + 100ug HEK293 cell lysate overexpressing GFP

  Lane 4: IP: Anti-GFP antibody (Magnetic beads) + 100ug HEK293 cell lysate overexpressing GFP

  Lane 5: HEK293 cell lysate overexpressing GFP - 10ug

  Lane 6: Unbound IP supernatant: anti-GFP antibody (Anti-GFP antibody ab290) + Protein A agarose + 100ug HEK293 cell lysate overexpressing GFP

  Lane 7: IP: anti-GFP antibody (Anti-GFP antibody ab290) + Protein A agarose+ 100ug HEK293 cell lysate overexpressing GFP

  Anti-GFP antibody (Magnetic beads) (Magnetic beads Anti-GFP VHH Single Domain antibody ab193983) was used to pull down GFP from HEK293 cell lysate overexpressing GFP. Anti-GFP antibody ab290 (Rabbit polyclonal to GFP - ChIP Grade) was used as a positive control. The gel was transferred to Nitrocellulose membrane and probed with mouse anti-GFP (Anti-GFP antibody [9F9.F9] ab1218) and anti-mouse HRP (ab97040).

  All lanes: Immunoprecipitation - Magnetic beads Anti-GFP VHH Single Domain antibody (Magnetic beads Anti-GFP VHH Single Domain antibody ab193983) at 1/500 dilution

  All lanes: HEK293 whole cell lysate - Overexpressing GFP at 10 µg

  Secondary

  All lanes: Immunoprecipitation - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/20000 dilution

  Observed band size: 48 kDa

  Exposure time: 30s

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-NOX2/gp91phox antibody [54.1] (Anti-NOX2/gp91phox antibody [54.1] ab80897) at 1 µg/mL

  Lane 1: Human liver tissue lysate - total protein (ab29889) at 10 µg

  Lane 2: HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

  Lane 3: Human lymph node tissue lysate - total protein (ab29871) at 10 µg

  Lane 4: MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 65 kDa

  Observed band size: 29 kDa, 35 kDa, 65 kDa

  Exposure time: 12min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  The band observed at 26 kDa could potentially be a cleaved form of NDUFS3 due to the presence of a 36 amino acid transit peptide.

  All lanes: Western blot - Anti-NDUFS3 antibody [17D95] (Anti-NDUFS3 antibody [17D95] ab14711) at 1 µg/mL

  Lane 1: Human brain tissue lysate - total protein (ab29466) at 10 µg

  Lane 2: Human heart tissue lysate - total protein (ab29431) at 10 µg

  Lane 3: Heart (Mouse) Tissue Lysate at 10 µg

  Lane 4: HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 30 kDa

  Observed band size: 26 kDa, 57 kDa

  Exposure time: 1min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-hnRNP A1 antibody [9H10] (Anti-hnRNP A1 antibody [9H10] ab5832) at 1 µg/mL

  Lane 1: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

  Lane 2: Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

  Lane 3: HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

  Lane 4: HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 38 kDa

  Observed band size: 37 kDa, 43 kDa

  Exposure time: 30s

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-PAX5 antibody [mAbcam52588] (Anti-PAX5 antibody [mAbcam52588] ab52588) at 1 µg/mL

  Lane 1: Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate (blocked with 1% milk) at 20 µg

  Lane 2: HEK293 (Human embryonic kidney cell line) Whole Cell Lysate (blocked with 1% Milk) at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 42 kDa

  Observed band size: 45 kDa, 50 kDa

  Exposure time: 20min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  This image was generated using the ascites version of the product.

  All lanes: Western blot - Anti-Syntaxin 6 antibody [3D10] - Golgi Membrane Marker (Anti-Syntaxin 6 antibody [3D10] - Golgi Membrane Marker ab12370) at 1 µg/mL

  Lane 1: Brain (Mouse) Tissue Lysate at 10 µg

  Lane 2: Brain (Rat) Tissue Lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 29 kDa

  Observed band size: 26 kDa, 30.6 kDa

  Exposure time: 8min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-pan ATPase Alpha antibody [M7-PB-E9] (Anti-pan ATPase Alpha antibody [M7-PB-E9] ab2871) at 1/500 dilution

  Lane 1: Human kidney tissue lysate - total protein (ab30203) at 20 µg

  Lane 2: Kidney (Mouse) Tissue Lysate at 20 µg

  Lane 3: Kidney (Rat) Tissue Lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 112 kDa

  Observed band size: 102 kDa, 28 kDa, 60 kDa

  Exposure time: 2min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-Superoxide Dismutase 3/EC-SOD antibody [4G11G6] (Anti-Superoxide Dismutase 3/EC-SOD antibody [4G11G6] ab80946) at 1 µg/mL

  All lanes: Lung (Mouse) Tissue Lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 26 kDa

  Observed band size: 25 kDa, 54 kDa

  Exposure time: 1min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-alpha Tubulin (acetyl K40) antibody [6-11B-1] (Anti-alpha Tubulin (acetyl K40) antibody [6-11B-1] ab24610) at 5 µg/mL

  Lane 1: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

  Lane 2: NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 20 µg

  Lane 3: PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 20 µg

  Lane 4: Brain (Mouse) Tissue Lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Performed under reducing conditions.

  Predicted band size: 50 kDa

  Observed band size: 140 kDa, 25 kDa, 35 kDa, 55 kDa

  Exposure time: 150s

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 overnight at 4°C. Antibody binding was detected using an anti-mouse HRP (ab97040), and visualised using ECL development solution ECL Substrate Kit (High Sensitivity) ab133406

  All lanes: Western blot - Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1 µg/mL

  Lane 1: HeLa (Human epithelial cell line from cervix adenocarcinoma cell line) whole cell lysate at 10 µg

  Lane 2: NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

  Lane 3: PC12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 36 kDa, 50 kDa

  Observed band size: 50 kDa

  Exposure time: 150s

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system under denaturing, reducing conditions. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. After transfer, the membrane was blocked for an hour in 3% milk before being incubated overnight at 4°C with mouse monoclonal [H4B4] to LAMP2 (Anti-LAMP2 antibody [H4B4] - Lysosome Marker ab25631; diluted 1:5000). Antibody binding was detected using peroxidase labelled goat anti-mouse IgG (ab97040; diluted 1:50000) for an hour at room temperature and visualised using ECL development solution.

  All lanes: Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (Anti-LAMP2 antibody [H4B4] - Lysosome Marker ab25631) at 1/500 dilution

  Lane 1: HeLa cell lysate at 20 µg

  Lane 2: Jurkat cell lysate at 20 µg

  Lane 3: Human liver lysate at 20 µg

  Lane 4: Human liver membrane fraction lysate at 20 µg

  Lane 5: Human skeletal muscle lysate at 20 µg

  Lane 6: Human brain lysate at 20 µg

  Lane 7: Human kidney lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution

  Performed under reducing conditions.

  Predicted band size: 45 kDa

  Observed band size: 100 kDa, 110 kDa, 120 kDa

  Exposure time: 20min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-Fyn antibody [FYN-01] (Anti-Fyn antibody [FYN-01] ab1881) at 1 µg/mL

  All lanes: Western blot - Recombinant human Fyn protein (Recombinant human Fyn protein ab84696) at 0.01 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 60 kDa

  Exposure time: 10s

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-Glucose Transporter GLUT2 antibody [5D1] (Anti-Glucose Transporter GLUT2 antibody [5D1] ab85715) at 1 µg/mL

  All lanes: Human pancreas tissue lysate - total protein (ab29816) at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 57 kDa

  Observed band size: 57 kDa

  Exposure time: 8min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  This image was generated using the ascites version of the product.

  All lanes: Western blot - Anti-HSD3B1 antibody [3C11-D4] (Anti-HSD3B1 antibody [3C11-D4] ab55268) at 1 µg/mL

  All lanes: Human placenta tissue lysate - total protein (ab29745) at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 42 kDa

  Observed band size: 42 kDa, 73 kDa

  Exposure time: 4min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  Anti-ETS1 antibody [mAbcam63302] ab63302 is predicted to react with both isoforms of ETS1. c-ETS-1A has a predicted molecular weight of 50-kDa, whereas c-ETS-1B has a predicted molecular weight of 41-kDa. The ETS1 protein sequence contains several phosphorylation sites (Swissprot), which might explain the banding pattern observed.

  All lanes: Western blot - Anti-ETS1 antibody [mAbcam63302] (Anti-ETS1 antibody [mAbcam63302] ab63302) at 10 µg/mL

  Lane 1: HepG2 (Human hepatocellular liver carcinoma cell line) Nuclear Lysate at 10 µg

  Lane 2: Jurkat nuclear extract lysate (ab14844) at 10 µg

  Lane 3: HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 10 µg

  Lane 4: MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 50 kDa

  Observed band size: 45 kDa, 55 kDa, 60 kDa

  Exposure time: 2min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-Alpha-synuclein antibody [LB 509] (Anti-Alpha-synuclein antibody [LB 509] ab27766) at 1/1000 dilution

  All lanes: Western blot - Recombinant Human Alpha-synuclein protein (Recombinant Human Alpha-synuclein protein ab51189) at 0.1 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 14 kDa

  Exposure time: 8min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  Anti-Amphiregulin antibody [MM0089-8K16] ab89119 recognizes the recombinant full length soluble Human Amphiregulin protein (Recombinant human Amphiregulin protein ab104355) which has an expected molecular weight of ~12 kDa.

  All lanes: Western blot - Anti-Amphiregulin antibody [MM0089-8K16] (Anti-Amphiregulin antibody [MM0089-8K16] ab89119) at 1/500 dilution

  All lanes: Western blot - Recombinant human Amphiregulin protein (Recombinant human Amphiregulin protein ab104355) at 0.1 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 28 kDa

  Exposure time: 8min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  The band observed at 99 kDa could potentially be a cleaved form of Metabotropic Glutamate Receptor 2/MGLUR2 due to the presence of an 18 amino acid signal peptide. Metabotropic Glutamate Receptor 2/MGLUR2 also contains a number of potential glycosylation sites (SwissProt) which may explain the fuzzy banding pattern observed.

  All lanes: Anti-Metabotropic Glutamate Receptor 2/MGLUR2 antibody [mG2Na-s] (ab15672) at 1 µg/mL

  All lanes: Brain (Rat) Tissue Lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Exposure time: 12min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

  All lanes: Western blot - Anti-SLC22A3/OCT3 antibody [mAbcam75402] (Anti-SLC22A3/OCT3 antibody [mAbcam75402] ab75402) at 5 µg/mL

  All lanes: Human prostate tissue lysate (cirrhosis) - total protein (ab30307) at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 61 kDa

  Observed band size: 61 kDa

  Exposure time: 16min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-NG2 antibody [132.38] (Anti-NG2 antibody [132.38] ab50009) at 1 µg/mL

  Lane 1: Rat brain tissue lysate at 20 µg

  Lane 2: Rat spinal cord tissue lysate at 20 µg

  Lane 3: Rat hippocampus tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 251 kDa

  Observed band size: 270 kDa, 31 kDa, 41 kDa

  Exposure time: 20min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-Lamin B2 antibody [LN43] (Anti-Lamin B2 antibody [LN43] ab8983) at 1 µg/mL

  Lane 1: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

  Lane 2: Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

  Lane 3: U2OS (Human osteosarcoma cell line) Whole Cell Lysate at 10 µg

  Lane 4: HL60 (Human promyelocytic leukemia cell line) Whole Cell Lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 68 kDa

  Observed band size: 37 kDa, 75 kDa

  Exposure time: 20min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  The binding pattern of Anti-GATA3 antibody [mAbcam77073] ab77073 in human thymus, lymph node, spleen and tonsil tissue lysates is similar to that of the polyclonal antibody to the same target (ab32858) in MOLT4, Jurkat, THP1 and Ramos whole cell lysates.

  All lanes: Western blot - Anti-GATA3 antibody [mAbcam77073] (Anti-GATA3 antibody [mAbcam77073] ab77073) at 5 µg/mL

  Lane 1: Human thymus tissue lysate - total protein (ab30146) at 20 µg

  Lane 2: Human lymph node tissue lysate - total protein (ab29871) at 20 µg

  Lane 3: Human spleen tissue lysate - total protein (ab29699) at 20 µg

  Lane 4: Tonsil (Human) Tissue Lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 47 kDa

  Observed band size: 55 kDa, 90 kDa

  Exposure time: 12min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-Cytochrome P450 1A2 antibody [d15 (16VII F10F12)] (Anti-Cytochrome P450 1A2 antibody [d15 (16VII F10F12)] ab22717) at 1 µg/mL

  Lane 1: Human liver tissue lysate - total protein (ab29889) at 10 µg

  Lane 2: Liver (Mouse) Tissue Lysate at 10 µg

  Lane 3: Liver (Rat) Tissue Lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 58 kDa

  Observed band size: 30 kDa, 48 kDa, 58 kDa

  Exposure time: 150s

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-hnRNP L antibody [4D11] (Anti-hnRNP L antibody [4D11] ab6106) at 1 µg/mL

  Lane 1: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

  Lane 2: Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

  Lane 3: MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

  Lane 4: HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 64 kDa

  Observed band size: 60 kDa, 64 kDa, 68 kDa

  Exposure time: 1min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  Muscarinic Acetylcholine Receptor 2/CM2 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

  All lanes: Western blot - Anti-Muscarinic Acetylcholine Receptor 2/CM2 antibody [31-1D1] (Anti-Muscarinic Acetylcholine Receptor 2/CM2 antibody [31-1D1] ab2805) at 1 µg/mL

  Lane 1: Human brain tissue lysate - total protein (ab29466) at 10 µg

  Lane 2: Human spinal cord tissue lysate - total protein (ab29188) at 10 µg

  Lane 3: Brain (Mouse) Tissue Lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 52 kDa

  Observed band size: 22 kDa, 40 kDa, 64 kDa

  Exposure time: 30s

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Anti-Cathepsin H antibody [3G8] (ab40699) at 1/1000 dilution

  All lanes: Western blot - Native human Cathepsin H protein (Native human Cathepsin H protein ab90385) at 0.01 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Exposure time: 1min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-Cytokeratin 18 antibody [RGE53] (Anti-Cytokeratin 18 antibody [RGE53] ab9217) at 5 µg/mL

  Lane 1: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

  Lane 2: MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 48 kDa

  Observed band size: 48 kDa

  Exposure time: 30s

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  Podoplanin protein is extensively glycosylated. This might explain the observation of a band at a higher molecular weight than predicted.

  All lanes: Western blot - Anti-Podoplanin / gp36 antibody [18H5] - BSA and Azide free (Anti-Podoplanin antibody [18H5] - BSA and Azide free ab10288) at 5 µg/mL

  Lane 1: Human placenta tissue lysate - total protein (ab29745) at 20 µg

  Lane 2: Human skeletal muscle tissue lysate - total protein (ab29330) at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 16 kDa

  Observed band size: 35 kDa

  Exposure time: 20min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-Cyclin A2 antibody [E23.1] (Anti-Cyclin A2 antibody [E23.1] ab38) at 5 µg/mL

  Lane 1: HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 20 µg

  Lane 2: U2OS (Human osteosarcoma cell line) Whole Cell Lysate at 20 µg

  Lane 3: PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 20 µg

  Lane 4: NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 49 kDa

  Observed band size: 50 kDa

  Exposure time: 20min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-CARD15/NOD2 antibody [2D9] (Anti-CARD15/NOD2 antibody [2D9] ab31488) at 2 µg/mL

  All lanes: HEK 293 overexpressing CARD15/NOD2 at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 115 kDa

  Observed band size: 115 kDa

  Exposure time: 150s

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  This image was generated using the ascites version of the product.

  All lanes: Western blot - Anti-HNF-4-alpha antibody [K9218] (Anti-HNF-4-alpha antibody [K9218] ab41898) at 1 µg/mL

  All lanes: HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 53 kDa

  Observed band size: 108 kDa, 37 kDa, 53 kDa

  Exposure time: 4min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-TIE2 antibody [Cl. 16] - BSA and Azide free (Anti-TIE2 antibody [Cl. 16] - BSA and Azide free ab24859) at 1 µg/mL

  All lanes: Western blot - Recombinant Human TIE2 protein (Fc Chimera) (Recombinant Human TIE2 protein (Fc Chimera) ab54435) at 0.01 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 125 kDa

  Exposure time: 2min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-pan Cadherin antibody [mAbcam22744] (Anti-pan Cadherin antibody [mAbcam22744] ab22744) at 1 µg/mL

  Lane 1: Heart (Rat) Tissue Lysate (blocked with 5% Milk) at 20 µg

  Lane 2: Heart (Mouse) Tissue Lysate (blocked with 5% Milk) at 20 µg

  Lane 3: Heart (Human) Tissue Lysate - adult normal tissue (blocked with 5% Milk) at 20 µg

  Lane 4: Heart (Rat) Tissue Lysate (blocked with 3% Milk) at 20 µg

  Lane 5: Heart (Mouse) Tissue Lysate (blocked with 3% Milk) at 20 µg

  Lane 6: Heart (Human) Tissue Lysate - adult normal tissue (blocked with 3% Milk) at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 125 kDa

  Observed band size: 125 kDa, 25 kDa, 58 kDa

  Exposure time: 8min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  Abcam recommends using milk (5%) as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
  

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (Anti-Cytochrome C antibody [7H8.2C12] ab13575).

  All lanes: Western blot - Anti-Cytochrome C antibody [7H8.2C12] (Anti-Cytochrome C antibody [7H8.2C12] ab13575) at 1 µg/mL

  Lane 1: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

  Lane 2: Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

  Lane 3: Human heart tissue lysate - total protein (ab29431) at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 11 kDa

  Observed band size: 14 kDa

  Exposure time: 3min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  We recommend using 5% milk in TBST as the blocking agent, decreasing to 2% milk in TBST during primary and secondary antibody incubation.
  

  Blots were developed with Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) secondary antibody

  All lanes: Western blot - Anti-HIF-1 alpha antibody [H1alpha67] (Anti-HIF-1 alpha antibody [H1alpha67] ab1) at 5 µg/mL

  Lane 1: Western blot - HeLa nuclear extract lysate (HeLa nuclear extract lysate ab150036) at 40 µg

  Lane 2: Western blot - Hela-DFO treated (0.5mM, 24h) Nuclear Lysate (Hela-DFO treated (0.5mM, 24h) Nuclear Lysate ab180880) at 40 µg

  Lane 3: HeLa nuclear control at 40 µg

  Lane 4: HeLa nuclear DFO treated at 40 µg

  Lane 5: Western blot - Recombinant Human HIF-1 alpha protein (Recombinant Human HIF-1 alpha protein ab154478) at 0.001 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

  Performed under reducing conditions.

  Predicted band size: 92 kDa

  Exposure time: 20min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker (Anti-ATPB antibody [4.3E8.D10] - Mitochondrial Marker ab5432) at 1 µg/mL

  All lanes: Human testis tissue lysate - total protein (ab30257) at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 56 kDa

  Observed band size: 57 kDa

  Exposure time: 16min

• 

  Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040)

  All lanes: Western blot - Anti-CDKN2A/p16INK4a antibody [DCS50.1] (Anti-CDKN2A/p16INK4a antibody [DCS50.1] ab16123) at 1 µg/mL

  Lane 1: HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

  Lane 2: HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 17 kDa

  Observed band size: 16 kDa, 37 kDa, 50 kDa

  Exposure time: 12min