Goat anti-rabbit IgG H&L (HRP) is a secondary antibody for chemiluminescent detection. Ideal for western blot. Suitable for ICC, IHC, western blot and ELISA.
- Cited in over 1090 publications
Application | Reactivity | Dilution info | Notes |
---|---|---|---|
Application IHC-P | Reactivity Reacts | Dilution info 1/200 - 1/5000 | Notes - |
Application ICC | Reactivity Reacts | Dilution info - | Notes - |
Application ELISA | Reactivity Reacts | Dilution info 1/10000 - 1/100000 | Notes - |
Application WB | Reactivity Reacts | Dilution info 1/2000 - 1/20000 | Notes Colorimetric: 1/5000 - 1/30000; Chemiluminescent: 1/10000 - 1/50000 |
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Ig gamma 1 chain C region, Ig gamma 3 chain C region, Ig gamma 4 chain C region, Ig gamma chain C region, Ig gamma-2 chain C region, Ig kappa-b4 chain C region, Ig kappa-b5 chain C region, Ig kappa-b9 chain C region, Ig lambda chain C region, Immunoglobin heavy constant gamma 1, Immunoglobulin G, K-BAS
Goat anti-rabbit IgG H&L (HRP) is a secondary antibody for chemiluminescent detection. Ideal for western blot. Suitable for ICC, IHC, western blot and ELISA.
- Cited in over 1090 publications
By immunoelectrophoresis and ELISA this antibody reacts specifically with Rabbit IgG and with light chains common to other Rabbit immunoglobulins. No antibody was detected against non-immunoglobulin serum proteins.
pH: 6.8 - 7.4
Preservative: 0.05% CMIT/MIT based preservative
Constituents: PBS, 0.2% BSA
This antibody was isolated by affinity chromatography using antigen coupled to agarose beads and conjugated to Horse Radish Peroxidase (HRP).
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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
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
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
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 43 kDa
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
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
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
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
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
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
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
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
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
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
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
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
This data was developed using Anti-SMAD1 + SMAD5 + SMAD9 (phospho S463 + S465 + S467) antibody [EP728(2)AY] ab76296, the same antibody clone in a different buffer formulation.
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
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 52 kDa
Exposure time: 7s
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
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
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
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
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
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
We recommend to use 1%SDS Hot lysis prepare method.
We are unsure how to define the extra bands.
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
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
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 60 kDa
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
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
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
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
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
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
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
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
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
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
All lanes: Western blot - Anti-Integrin alpha V antibody [EPR16800] (Anti-Integrin alpha V antibody [EPR16800] ab179475) at 1/1000 dilution
Lane 1: Untreated HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate. at 15 µg
Lane 2: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate treated with PNGase F. at 15 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 116 kDa
Observed band size: 116.135 kDa
Exposure time: 20s
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
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
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
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Observed band size: 85 kDa
Exposure time: 100s
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
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 47 kDa
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
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
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
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 63 kDa
Exposure time: 30s
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
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
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
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
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
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
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
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
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
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
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
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
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)
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Exposure time: 180s
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
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 42 kDa
Exposure time: 1min
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
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
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