Goat Anti-Rabbit IgG H&L (HRP)

41 product images

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-RAB29 (phospho T71) antibody [MJF-R24-17-1] (Anti-RAB29 (phospho T71) antibody [MJF-R24-17-1] ab241062) at 1/5000 dilution

  Lane 1: HEK-293 (human embryonic kidney) over-expressing HA-tagged RAB29 and LRRK2 [Y1699C], whole cell lysate at 15 µg

  Lane 2: MLi-2 (100nM for 60min) treated HEK-293 over-expressing HA-tagged RAB29 and LRRK2 [Y1699C], whole cell lysate at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 23 kDa

  Observed band size: 23 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and dilution buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-Paxillin (phospho Y118) antibody [EPR1903] (Anti-Paxillin (phospho Y118) antibody [EPR1903] ab109547) at 1/1000 dilution

  Lane 1: Jurkat (human T cell leukemia T lymphocyte) whole cell lysate at 10 µg

  Lane 2: Jurkat treated with 50mM pervanadate for 5 min whole cell lysate at 10 µg

  Lane 3: Jurkat treated with 50mM pervanadate for 5 min whole cell lysate, then the membrane was incubated with alkaline phosphatase at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 65 kDa

  Observed band size: 65 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and diluting buffer: 5% NFDM/TBST.

  All lanes: Western blot - Anti-p53 (phospho S392) antibody [EP155Y] (Anti-p53 (phospho S392) antibody [EP155Y] ab33889) at 1/1000 dilution

  Lane 1: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates at 15 µg

  Lane 2: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates, then the membrane was incubated with alkaline phosphatase at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 43 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking/Diluting Buffer and concentration: 5% NFDM /TBST

  All lanes: Western blot - Anti-ATF2 (phospho T71) antibody [E268] (Anti-ATF2 (phospho T71) antibody [E268] ab32019) at 0.526 µg/mL

  Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) grown in serum free media for overnight, whole cell lysate at 20 µg

  Lane 2: HeLa grown in serum free media for overnight, then treated with 250ng/ml Anisomycin for 30min, whole cell lysate at 20 µg

  Lane 3: HeLa grown in serum free media for overnight, then treated with 250ng/ml Anisomycin for 30min whole cell lysate, then the membrane was incubated with alkaline phosphatase at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 0.05 µg/mL

  Predicted band size: 55 kDa

  Observed band size: 70 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  All lanes: Western blot - Anti-mTOR (phospho S2448) antibody [EPR426(2)] (Anti-mTOR (phospho S2448) antibody [EPR426(2)] ab109268) at 1/2000 dilution

  Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 10 µg

  Lane 2: HeLa grown in serum-free media overnight, then treated with 200nM PMA for 4 hours whole cell lysate at 10 µg

  Lane 3: HeLa grown in serum-free media overnight, then treated with 200nM PMA for 4 hours whole cell lysate. Then the membrane was incubated with alkaline phosphatase at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 289 kDa

  Observed band size: 289 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and dilution buffer: 5% NFDM/TBST.
  

  Exposure times:

  Lanes 1-6: 3 seconds;
  Lane 7: 7 seconds.

  The molecular weight observed is consistent with what has been described in the literature (PMID:10514407).

  All lanes: Western blot - Anti-PTEN antibody [EPR22636-122] (Anti-PTEN antibody [EPR22636-122] ab267787) at 1/1000 dilution

  Lane 1: C6 (rat glial tumor glial cell), whole cell lysate at 10 µg

  Lane 2: RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 10 µg

  Lane 3: PC-12 (rat adrenal gland pheochromocytoma), whole cell lysate at 10 µg

  Lane 4: NIH/3T3 (mouse embryonic fibroblast), whole cell lysate at 10 µg

  Lane 5: Mouse kidney tissue lysate at 10 µg

  Lane 6: Mouse spleen tissue lysate at 10 µg

  Lane 7: Rat lung tissue lysate at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 47 kDa

  Observed band size: 54 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Diluting/Blocking buffer and concentration: 5% NFDM/TBST
  

  It detects an 70kDa extra band in some cell lines.

  All lanes: Western blot - Anti-Zic2 antibody [EPR7790] (Anti-Zic2 antibody [EPR7790] ab150404) at 1/2000 dilution

  Lane 1: Human fetal brain tissue lysate at 20 µg

  Lane 2: SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: C6 (Rat glial tumor glial cell) whole cell lysate at 20 µg

  Lane 4: Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 25 kDa, 55 kDa

  Observed band size: 28 kDa, 56 kDa

  Exposure time: 3min

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  All lanes: Western blot - Anti-TUBA1B antibody [EPR1333] - Loading Control (Anti-TUBA1B antibody [EPR1333] - Loading Control ab108629) at 1/100000 dilution

  Lane 1: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates at 20 µg

  Lane 2: Mouse brain lysates at 20 µg

  Lane 3: Mouse kidney lysates at 20 µg

  Lane 4: Rat brain lysates at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 50 kDa

  Observed band size: 50 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-Actin antibody [EPR19353-16] - Loading Control ab216070, the same antibody clone in a different buffer formulation.
  

  Blocking and dilution buffer: 5% NFDM/TBST.

  Human Gamma actin (ACTG1) contains aa2-182 with His-tag. Human Beta actin (ACTB) contains aa1-100 with GST and His tag. Human Gamma-2 actin (ACTG2) contains aa1-200 with His-tag. Human Alpha-cardiac actin (ACTC) contains aa1-200 with His-tag. Human Alpha-actin-2 (ACTA2) contains aa3-253 with His-tag. Human Alpha-actin-1 (ACTA1) contains aa3-200 with His-tag.

  All lanes: Western blot - Anti-Actin antibody [EPR19353-16] - Loading Control (Anti-Actin antibody [EPR19353-16] - Loading Control ab216070) at 1/1000 dilution

  Lane 1: Human Gamma actin (ACTG1) recombinant protein at 0.01 µg

  Lane 2: Human Beta actin (ACTB) recombinant protein at 0.02 µg

  Lane 3: Human Gamma-2 actin (ACTG2) recombinant protein at 0.02 µg

  Lane 4: Human Alpha-cardiac actin (ACTC) recombinant protein at 0.02 µg

  Lane 5: Human Alpha-actin-2 (ACTA2) recombinant protein at 0.01 µg

  Lane 6: Human Alpha-actin-1 (ACTA1) recombinant protein at 0.01 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

  Predicted band size: 42 kDa

  Exposure time: 1min

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-alpha Actinin 4 antibody [EPR2533(2)] - Loading Control ab108198, the same antibody clone in a different buffer formulation.

  All lanes: Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] - Loading Control (Anti-alpha Actinin 4 antibody [EPR2533(2)] - Loading Control ab108198) at 1/10000 dilution

  All lanes: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 105 kDa

  Observed band size: 105 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking/Dilution buffer: 5% NFDM/TBST.
  

  The MW observed is consistent with the literature (PMID 10704451; PMID 1384988).

  Negative control: liver, brain tissue (PMID 2578885, PMID 25288117).

  All lanes: Western blot - Anti-Myelin Protein Zero antibody [EPR20383] (Anti-Myelin Protein Zero antibody [EPR20383] ab183868) at 1/5000 dilution

  Lane 1: Mouse sciatic nerve lysate at 4 µg

  Lane 2: Mouse liver lysate at 4 µg

  Lane 3: Mouse brain lysate at 4 µg

  Lane 4: Rat sciatic nerve lysate at 4 µg

  Lane 5: Rat liver lysate at 4 µg

  Lane 6: Rat brain lysate at 4 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

  Predicted band size: 28 kDa

  Observed band size: 26-28 kDa

  Exposure time: 1s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [EP728(2)AY] (Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [EP728(2)AY] ab76296) at 1/1000 dilution

  Lane 1: Untreated C6 (Rat glial tumor glial cell) whole cell lysate at 15 µg

  Lane 2: C6 treated with 10ng/ml TGF beta 3 and 50µM MG-132 for 30 min whole cell lysate at 15 µg

  Lane 3: C6 treated with 10ng/ml TGF beta 3 and 50µM MG-132 for 30 min whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Observed band size: 52 kDa

  Exposure time: 7s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR19399] ab197072, the same antibody clone in a different buffer formulation.
  

  Blocking and dilution buffer: 5% NFDM/TBST.

  The observed MW is higher than the predicted one due to the glycosylation. The expression pattern is consistent with the published papers (PMID: 23115189 and 20064930).

  All lanes: Western blot - Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR19399] (Anti-TrkA (phospho Y674)+TrkB (phospho Y706)+TrkC (phospho Y709) antibody [EPR19399] ab197072) at 1/1000 dilution

  Lane 1: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with TrkB (wild type) with Myc-tag at 20 µg

  Lane 2: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with TrkB (wild type) with Myc-tag starved for 4 hours, then treated with 50 ng/ml BDNF for 10 minutes at 20 µg

  Lane 3: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with TrkB (phospho Y706A mutant) with Myc-tag at 20 µg

  Lane 4: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate transfected with TrkB (phospho Y706A mutant) with Myc-tag starved for 4 hours, then treated with 50 ng/ml BDNF for 10 minutes at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

  Predicted band size: 92 kDa

  Observed band size: 140 kDa

  Exposure time: 1s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and diluting buffer: 5% NFDM/TBST

  All lanes: Western blot - Anti-ATP5A antibody [EPR13030(B)] - Mitochondrial Marker (Anti-ATP5A antibody [EPR13030(B)] - Mitochondrial Marker ab176569) at 0.01 µg/mL

  Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

  Lane 2: Mouse brain lysates at 15 µg

  Lane 3: Rat brain lysates at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 60 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  All lanes: Western blot - Anti-ACTN3 antibody [EP2531Y] - Loading Control (Anti-ACTN3 antibody [EP2531Y] - Loading Control ab68204) at 1/20000 dilution

  Lane 1: Mouse skeletal muscle lysates at 15 µg

  Lane 2: Rat skeletal muscle lysates at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 103 kDa

  Observed band size: 100 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-NR1D1 + NR1D2 / RVR antibody [EPR10376] (Anti-NR1D1 + NR1D2 / RVR antibody [EPR10376] ab174309) at 1/1000 dilution

  Lane 1: GST-tagged human full length recombinant NR1D1 protein at 0.015 µg

  Lane 2: GST-tagged human full length recombinant NR1D2 protein at 0.015 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Observed band size: 85 kDa

  Exposure time: 100s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  All lanes: Western blot - Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - Neuronal Marker (Anti-ENO1 + ENO2 + ENO3 antibody [EPR3377] - Neuronal Marker ab79757) at 1/1000 dilution

  Lane 1: SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 3: Y79 (Human retinoblastoma retinoblastoma) whole cell lysate at 20 µg

  Lane 4: Mouse brain lysate at 20 µg

  Lane 5: Rat brain lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 47 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-Transferrin antibody [EPR18240-29] - Serum Loading Control ab278498, the same antibody clone in a different buffer formulation.
  

  Blocking and diluting buffer and concentration: 5% NFDM/TBST

  Exposure time: Lane 1-3: 3.25 seconds; Lane 4: 70 seconds

  All lanes: Western blot - Anti-Transferrin antibody [EPR18240-29] - Serum Loading Control (Anti-Transferrin antibody [EPR18240-29] - Serum Loading Control ab278498) at 1/1000 dilution

  Lane 1: Mouse liver tissue lysate at 20 µg

  Lane 2: Mouse kidney tissue lysate at 20 µg

  Lane 3: Mouse testis tissue lysate at 20 µg

  Lane 4: Hepa1-6 (mouse hepatoma epithelial cell), whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

  Predicted band size: 77 kDa

  Observed band size: 76 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-NAK/TBK1 (phospho S172) antibody [EPR2867(2)] (Anti-NAK/TBK1 (phospho S172) antibody [EPR2867(2)] ab109272) at 1/5000 dilution

  Lane 1: Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 2: HeLa was starved overnight, then treated with 100nM Calyculin A for 30 min whole cell lysate at 15 µg

  Lane 3: HeLa was starved overnight, then treated with 100nM Calyculin A for 30 min whole cell lysate, then the membrane treated with Phosphatase for 1 hour

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 84 kDa

  Observed band size: 84 kDa

  Exposure time: 37s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-CD81 antibody [EPR4244] (Anti-CD81 antibody [EPR4244] ab109201) at 1/1000 dilution

  Lane 1: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

  Lane 2: C2C12 (Mouse myoblasts myoblast) whole cell lysate at 20 µg

  Lane 3: Mouse brain tissue lysate at 20 µg

  Lane 4: Mouse heart tissue lysate at 20 µg

  Lane 5: Huh7 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 6: HL-60 (Human acute promyelocytic leukemia promyeloblast) whole cell lysate at 20 µg

  Lane 7: HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 8: LNCaP (Human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 9: U-87 MG (Human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

  Lane 10: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 11: A549 (Human lung carcinoma epithelial cell) whole cell lysate at 20 µg

  Lane 12: 293T (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 25 kDa

  Observed band size: 25 kDa

  Exposure time: 60s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking/Diluting buffer 5% NFDM/TBST

  All lanes: Western blot - Anti-Caveolin-1 (phospho Y14) antibody [EPR2288Y] - Caveolae Marker (Anti-Caveolin-1 (phospho Y14) antibody [EPR2288Y] - Caveolae Marker ab75876) at 1/1000 dilution

  Lane 1: Untreated A673 (Human muscle Ewing's Sarcoma) whole cell lysates at 15 µg

  Lane 2: A673 (Human muscle Ewing's Sarcoma) treated with insulin, whole cell lysates at 15 µg

  Lane 3: A673 (Human muscle Ewing's Sarcoma) treated with insulin, whole cell lysates. Then the membrane was incubated with phosphatase. at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 20 kDa

  Observed band size: 22 kDa

  Exposure time: 30s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-Myoglobin antibody [EP3081Y] (Anti-Myoglobin antibody [EP3081Y] ab77232) at 1/5000 dilution

  Lane 1: Human muscle lysate at 20 µg

  Lane 2: Mouse muscle lysate at 20 µg

  Lane 3: Rat muscle lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 17 kDa

  Observed band size: 17 kDa

  Exposure time: 1s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-PFKM + PFKL antibody [EPR11904] (Anti-PFKM + PFKL antibody [EPR11904] ab181064) at 1/1000 dilution

  Lane 1: GST tagged Recombinant Human PFKL protein (Full length, 116 KDa)

  Lane 2: DDK tagged Recombinant Human PFKM protein (Full length, 85 KDa)

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Exposure time: 180s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-RAGE antibody [EPR12206] ab172473, the same antibody clone in a different buffer formulation.
  

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  Exposure time:

  Lanes 1-6: 3 second
  Lanes 7-11: 20 seconds

  The expression profile and molecular mass observed is consistent with what has been described in the literature (PMID: 16315007; 18355449; 18245812).

  All lanes: Western blot - Anti-RAGE antibody [EPR12206] (Anti-RAGE antibody [EPR12206] ab172473) at 1/1000 dilution

  Lane 1: Mouse lung lysates at 20 µg

  Lane 2: Mouse brain lysates at 20 µg

  Lane 3: Mouse kidney lysates at 20 µg

  Lane 4: Mouse heart lysates at 20 µg

  Lane 5: Mouse liver lysates at 20 µg

  Lane 6: Mouse spleen lysates at 20 µg

  Lane 7: Rat lung lysates at 20 µg

  Lane 8: Rat brain lysates at 20 µg

  Lane 9: Rat kidney lysates at 20 µg

  Lane 10: Rat heart lysates at 20 µg

  Lane 11: Rat spleen lysates at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 42 kDa

  Observed band size: 43 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Cdc6 (phospho S54) antibody [EPR759Y] ab75809).

  All lanes: Western blot - Anti-Cdc6 (phospho S54) antibody [EPR759Y] - BSA and Azide free (Anti-Cdc6 (phospho S54) antibody [EPR759Y] - BSA and Azide free ab247402) at 1/5000 dilution

  Lane 1: Hela (human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg

  Lane 2: Hela (human cervix adenocarcinoma epithelial cell) treated with Hydroxyurea (3mM, 20 h) whole cell lysate at 15 µg

  Lane 3: Hela (human cervix adenocarcinoma epithelial cell) treated with Hydroxyurea (3mM, 20 h) whole cell lysate. Then the membrane was incubated with phosphatase at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 63 kDa

  Exposure time: 30s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-beta Catenin (phospho S37) antibody [EP742(2)Y] ab75777, the same antibody clone in a different buffer formulation.

  All lanes: Western blot - Anti-beta Catenin (phospho S37) antibody [EP742(2)Y] - BSA and Azide free (Anti-beta Catenin (phospho S37) antibody [EP742(2)Y] - BSA and Azide free ab247399) at 1/5000 dilution

  Lane 1: HT-1080 (Human fibrosarcoma epithelial cell) whole cell lysate at 15 µg

  Lane 2: HT-1080 (Human fibrosarcoma epithelial cell) whole cell lysate treated with MG-132 at 15 µg

  Lane 3: HT-1080 (Human fibrosarcoma epithelial cell) whole cell lysate treated with MG-132 then the membrane was incubated with phosphatase at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 85 kDa

  Observed band size: 100 kDa

  Exposure time: 30s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  All lanes: Western blot - Anti-p53 (phospho S37) antibody [EPR17353] (Anti-p53 (phospho S37) antibody [EPR17353] ab182164) at 1/1000 dilution

  Lane 1: A431 whole cell lysate at 10 µg

  Lane 2: A431 treated with 1μg/ml doxorubicin for 24 hours whole cell lysate at 10 µg

  Lane 3: A431 treated with 1μg/ml doxorubicin for 24 hours whole cell lysate, the membrane was incubated with alkaline phosphatase at 10 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 43 kDa

  Observed band size: 53 kDa

  Exposure time: 15s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  All lanes: Western blot - Anti-RPS6 (phospho S235) antibody [EP1338(2)Y] - Ribosome Marker (Anti-RPS6 (phospho S235) antibody [EP1338(2)Y] - Ribosome Marker ab80158) at 1/1000 dilution

  Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) serum starved for 24 h. Whole cell lysates at 15 µg

  Lane 2: HeLa (Human cervix adenocarcinoma epithelial cell) serum starved for 24 h, then 20% FBS incubated for 90 minutes. Whole cell lysates at 15 µg

  Lane 3: HeLa (Human cervix adenocarcinoma epithelial cell) serum starved for 24 h, then 20% FBS incubated for 90 minutes. Whole cell lysates. Then the membrane was incubated with phosphatase at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 29 kDa

  Observed band size: 30 kDa

  Exposure time: 1min

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  The expression profile and molecular mass observed is consistent with what has been described in the literature (PMID:16315007; PMID:18355449; PMID:18245812).
  

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-RAGE antibody [EPR21171] (Anti-RAGE antibody [EPR21171] ab216329) at 1/2000 dilution

  Lane 1: Mouse lung lysates at 20 µg

  Lane 2: Mouse brain lysates at 20 µg

  Lane 3: Mouse kidney lysates at 20 µg

  Lane 4: Mouse heart lysates at 20 µg

  Lane 5: Mouse liver lysates at 20 µg

  Lane 6: Mouse spleen lysates at 20 µg

  Lane 7: Rat lung lysates at 20 µg

  Lane 8: Rat brain lysates at 20 µg

  Lane 9: Rat kidney lysates at 20 µg

  Lane 10: Rat heart lysates at 20 µg

  Lane 11: Rat spleen lysates at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 42 kDa

  Observed band size: 43 kDa

  Exposure time: 3s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-PON2 antibody [EPR15295-58] (Anti-PON2 antibody [EPR15295-58] ab183718) at 1/1000 dilution

  Lane 1: Mouse brain tissue lysate at 20 µg

  Lane 2: Mouse heart tissue lysate at 20 µg

  Lane 3: Mouse kidney tissue lysate at 20 µg

  Lane 4: Mouse spleen tissue lysate at 20 µg

  Lane 5: Rat brain tissue lysate at 20 µg

  Lane 6: Rat heart tissue lysate at 20 µg

  Lane 7: Rat kidney tissue lysate at 20 µg

  Lane 8: Rat spleen tissue lysate at 20 µg

  Lane 9: C6 (Rat glial tumor glial cell) whole cell lysate at 20 µg

  Lane 10: Raw264.7 (Mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

  Lane 11: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

  Lane 12: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 39 kDa

  Observed band size: 39 kDa

  Exposure time: 30s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  p53 K382 acetylation is responsed to cell stress such as DNA damage caused by ultraviolet or ionizing radiation as was described in PMID 9744860 and 11250899

  All lanes: Western blot - Anti-p53 (acetyl K382) antibody [EPR358(2)] (Anti-p53 (acetyl K382) antibody [EPR358(2)] ab75754) at 1/1000 dilution

  Lane 1: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 15 µg

  Lane 2: HepG2 (Human hepatocellular carcinoma epithelial cell) was starved overnight, then treatment with 30ug/ml etoposide for 8 hours followed by 500ng/ml Trichostatin A for 4 hours whole cell lysates at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 43 kDa

  Observed band size: 53 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  The doublets are reported by PMID: 9339377, 28817674 and 10051604.
  

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-PEX19 antibody [EPR9266(B)] (Anti-PEX19 antibody [EPR9266(B)] ab137072) at 1/1000 dilution

  Lane 1: Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates at 20 µg

  Lane 2: MOLT-4 (Human lymphoblastic leukemia T lymphoblast) whole cell lysates at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 33 kDa

  Observed band size: 35 kDa, 40 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Nanog is highly expressed in cancer stem cells. Although some papers support the expression in undifferentiated cancer cell lines, such as HeLa (PMID: 22337995, 28092370), MDA-MB-231 (PMID: 28401007, 25919570), HepG2 (PMID: 29477378), Huh7 (PMID: 26919045), HCT 116 (PMID: 25249558, 28092370) and PANC-1 (PMID: 28703793, 25846752), Anti-Nanog antibody [EPR2027(2)] ab109250 can't detect the target band in these cell lines, even at the dilution of 1:200.
  

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-Nanog antibody [EPR2027(2)] (Anti-Nanog antibody [EPR2027(2)] ab109250) at 1/2000 dilution

  Lane 1: NCCIT (Human pluripotent embryonic carcinoma epithelial cell) whole cell lysates at 20 µg

  Lane 2: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

  Lane 3: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates at 20 µg

  Lane 4: MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 20 µg

  Lane 5: HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 20 µg

  Lane 6: Huh7 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 20 µg

  Lane 7: HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysates at 20 µg

  Lane 8: PANC-1 (Human pancreatic epithelioid carcinoma epithelial cell) whole cell lysates at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 34 kDa

  Observed band size: 37 kDa

  Exposure time: 30s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] (Anti-JNK1 + JNK2 + JNK3 (phospho T183+T183+T221) antibody [EPR5693] ab124956) at 1/1000 dilution

  Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) Whole cell lysates at 15 µg

  Lane 2: HeLa (Human cervix adenocarcinoma epithelial cell) treated with 20J/m2 UV-C then recovery for 1 hour whole cell lysates at 15 µg

  Lane 3: HeLa (Human cervix adenocarcinoma epithelial cell) treated with 20J/m2 UV-C then recovery for 1 hour whole cell lysates. Then the membrane was incubated with alkaline phosphatase at 15 µg

  Lane 4: HeLa (Human cervix adenocarcinoma epithelial cell) treated with 20J/m2 UV-C then recovery for 1 hour whole cell lysates. Then the membrane was incubated with lambda phosphatase at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Observed band size: 46 kDa, 54 kDa

  Exposure time: 30s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-Integrin alpha V antibody [EPR16800] (Anti-Integrin alpha V antibody [EPR16800] ab179475) at 1/1000 dilution

  All lanes: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate treated with PNGase F. at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 116 kDa

  Observed band size: 116 kDa, 135 kDa

  Exposure time: 20s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  All lanes: Western blot - Anti-TWEAKR/FN14 antibody [EPR3179] (Anti-TWEAKR/FN14 antibody [EPR3179] ab109365) at 1/1000 dilution

  Lane 1: NIH/3T3 (mouse embryo) whole cell lysate at 20 µg

  Lane 2: L6 (rat skeletal muscle) whole cell lysate at 20 µg

  Lane 3: C2C12 (mouse muscle) whole cell lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 14 kDa

  Observed band size: 14-17 kDa

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-CD36 antibody (Anti-CD36 antibody ab124515) at 1/5000 dilution

  Lane 1: Mouse platelet tissue lysate at 20 µg

  Lane 2: Mouse adipose tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 53 kDa

  Observed band size: 75 kDa

  Exposure time: 90s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-Vitronectin/S-Protein antibody [EP873Y] (Anti-Vitronectin/S-Protein antibody [EP873Y] ab45139) at 1/2000 dilution

  All lanes: Human serum tissue lysate at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 54 kDa

  Observed band size: 75 kDa

  Exposure time: 30s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-GLP-1R antibody [EPR21819] (Anti-GLP-1R antibody [EPR21819] ab218532) at 1/1000 dilution

  Lane 1: Beta-TC-6 (Mouse pancreas insulinoma beta cell) whole cell lysate boiled. at 20 µg

  Lane 2: Beta-TC-6 (Mouse pancreas insulinoma beta cell) whole cell lysate unboiled. at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

  Predicted band size: 53 kDa

  Observed band size: 46.66 kDa

  Exposure time: 20s

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  We recommend to use 1%SDS Hot lysis prepare method.
  

  We are unsure how to define the extra bands.

  Blocking and diluting buffer and concentration: 5% NFDM/TBST.

  All lanes: Western blot - Anti-HIRA/HIR antibody [EPR7416] (Anti-HIRA/HIR antibody [EPR7416] ab129169) at 1/1000 dilution

  Lane 1: HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method at 20 µg

  Lane 2: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates prepared in 1%SDS Hot lysis method at 20 µg

  Lane 3: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates prepared in RIPA lysis method at 20 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 112 kDa

  Observed band size: 112 kDa

  Exposure time: 3min

• 

  Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

  This data was developed using Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298, the same antibody clone in a different buffer formulation.

  All lanes: Western blot - Anti-Rb (phospho T356) antibody [EPR2153AY] (Anti-Rb (phospho T356) antibody [EPR2153AY] ab76298) at 1/4000 dilution

  Lane 1: K562 (Human chronic myelogenous leukemia cell line from bone marrow) 2 day serum starved whole cell lysate at 15 µg

  Lane 2: K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate at 15 µg

  Lane 3: K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate, membrane incubated with phosphatase at 15 µg

  Secondary

  All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

  Predicted band size: 106 kDa

  Observed band size: 106 kDa

  Exposure time: 1min