VeriBlot for IP Detection Reagents are immunoblotting reagents that enable the trouble-free detection of immunoblotted target protein bands, without interference from denatured IgG.
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- Cell reports 40:1111072022Alphaherpesvirus US3 protein-mediated inhibition of the m6A mRNA methyltransferase complex.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRobert J J Jansens,Ruth Verhamme,Aashiq H Mirza,Anthony Olarerin-George,Cliff Van Waesberghe,Samie R Jaffrey,Herman W FavoreelPubMed 35858564
- The EMBO journal 40:e1062142021BNIP3 promotes HIF-1α-driven melanoma growth by curbing intracellular iron homeostasis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMónica Vara-Pérez,Matteo Rossi,Chris Van den Haute,Hannelore Maes,Maria Livia Sassano,Vivek Venkataramani,Bernhard Michalke,Erminia Romano,Kristine Rillaerts,Abhishek D Garg,Corentin Schepkens,Francesca M Bosisio,Jasper Wouters,Ana Isabel Oliveira,Peter Vangheluwe,Wim Annaert,Johannes V Swinnen,Jean Marie Colet,Joost J van den Oord,Sarah-Maria Fendt,Massimiliano Mazzone,Patrizia AgostinisPubMed 33932034
- Scientific reports 11:72562021A novel proteomics approach to epigenetic profiling of circulating nucleosomes.Applications:Unspecified applicationReactive species:Unspecified reactive speciesPriscilla Van den Ackerveken,Alison Lobbens,Jean-Valery Turatsinze,Victor Solis-Mezarino,Moritz Völker-Albert,Axel Imhof,Marielle HerzogPubMed 33790358
- Viruses 13:2021Discovery of RSV-Induced BRD4 Protein Interactions Using Native Immunoprecipitation and Parallel Accumulation-Serial Fragmentation (PASEF) Mass Spectrometry.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMorgan Mann,David S Roberts,Yanlong Zhu,Yi Li,Jia Zhou,Ying Ge,Allan R BrasierPubMed 33799525
- Frontiers in oncology 11:6251692021NEDD4 Induces K48-Linked Degradative Ubiquitination of Hepatitis B Virus X Protein and Inhibits HBV-Associated HCC Progression.Applications:Unspecified applicationReactive species:Unspecified reactive speciesTao Wan,Zhao Lei,Biao Tu,Tianyin Wang,Jiale Wang,Feizhou HuangPubMed 33767993
- Communications biology 4:3182021Bmp8a is an essential positive regulator of antiviral immunity in zebrafish.Applications:Unspecified applicationReactive species:Unspecified reactive speciesShenjie Zhong,Haoyi Li,Yun-Sheng Wang,Ying Wang,Guangdong Ji,Hong-Yan Li,Shicui Zhang,Zhenhui LiuPubMed 33750893
- Scientific reports 11:20972021Loss of Parkinson's susceptibility gene LRRK2 promotes carcinogen-induced lung tumorigenesis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesChandra Lebovitz et. al.PubMed 33483550
- Molecular brain 14:142021Schizophrenia-associated dysbindin modulates axonal mitochondrial movement in cooperation with p150.Applications:Unspecified applicationReactive species:Unspecified reactive speciesBo Kyoung Suh et. al.PubMed 33461576
- The Biochemical journal :2021Temporal modulation of the NF-κB RelA network in response to different types of DNA damage.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAmy E Campbell et. al.PubMed 33438746
- JCI insight :2021CMTM6 drives cisplatin resistance by regulating Wnt signaling through ENO-1/AKT/GSK3β axis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesPallavi Mohapatra,Omprakash Shriwas,Sibasish Mohanty,Arup Ghosh,Shuchi Smita,Sandeep Rai Kaushik,Rakesh Arya,Rachna Rath,Saroj Das Majumdar,Dillip Kumar Muduly,Sunil Raghav,Ranjan K Nanda,Rupesh DashPubMed 33434185
Key facts
- Applications
WB
- Form
Liquid
Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application WB | Reactivity Reacts | Dilution info 1/40.00000 - 1/4000.00000 | Notes The dilution will depend on the sensitivity of the HRP substrate. The dilution range recommended is 1:40 - 1:4000. Based on a 1:200 dilution (25 μL) in 5 ml milk researchers can perform 20 western blots. This product is recommended for the western blot detection of IP samples. Make sure the lysates are reduced and denatured completely. |
Recommended products
VeriBlot for IP Detection Reagents are immunoblotting reagents that enable the trouble-free detection of immunoblotted target protein bands, without interference from denatured IgG.
Key facts
- Applications
WB
- Form
Liquid
- Storage buffer
Constituents: 1% MOPS
Storage
- Shipped at conditions
Blue Ice
- Appropriate long-term storage conditions
+4°C
Notes
VeriBlot for IP Detection Reagents are immunoblotting reagents that enable the trouble-free detection of immunoblotted target protein bands, without interference from denatured IgG. This allows to detect the (co-)immunoprecipitated protein without masking by the IgG heavy (50 kDa) and light chains (25 kDa). In general, this interference tends to originate from secondary antibodies which recognize primary antibodies released with the antigen during the immunoprecipitation procedure or endogenous IgGs from the lysate itself. VeriBlot for IP detection reagents only recognize native (non-reduced) antibodies and therefore the detection of heavy and light chains is highly minimized, if the immunoprecipitate is fully reduced.
Number of blots:
At least 20 (based on a 1:200 dilution in 5 ml milk).
Important protocol notes (This information is available in Chinese here)
1. The VeriBlot for IP Detection Reagent (HRP) detects the following IgG polyclonal and monoclonal antibodies:
| Species | Monoclonal Isotype(s) |
| Bovine | IgG2 |
| Goat | IgG2 |
| Human | IgG1, IgG2, IgG4 |
| Mouse | IgG2a, IgG2b, IgG3 Note: If using mouse IgG1, perform a dot blot to determine compatibility. VeriBlot for IP Detection Reagent (HRP) might not detect mouse IgG1. |
| Rat | IgG2C |
| Rabbit | Total IgG |
| Sheep | IgG2 |
2. The VeriBlot for IP Detection Reagent (HRP) preferentially detects the non-reduced form over the reduced, SDS-denatured forms.
3. IP sample should be completely reduced/denatured before loaded onto a western blot. Boil samples for 5-10 minutes in SDS sample buffer with a increase in SDS amount if required.
4. Milk should be used as the blocking protein for the immunoblot.
Note: If denatured and blotted IgG are not clearly detected, the following steps may be used to increase the amount of denatured IgG in the sample:
- Increase the concentration of reducing agent
- Boil sample to aid in reduction of IgG disulfide bonds
- Use dentaturing electrophoresis conditions
A full troubleshooting guide is available here.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
31 product images
This image is courtesy of an anonymous customer review.Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Anti-IRAK2 antibody ab6148 Immunoprecipitating IRAK2 in human HEK293 whole cell lysate. 1000μg of cell lysate was incubated with primary antibody (1 μg/mg) and matrix (Protein G) for 16 hours at 4°C. For western blotting a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) (1/10000) was used to confirm successful immunoprecipation.
Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Lane 1 (input): HeLa (human cervix adenocarcinoma) treated with 250ng/ml anisomycin for 30min whole cell lysate 10μg.
Lane 2 (+): HeLa treated with 250ng/ml anisomycin for 30min whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-ATF2 (phospho T71) antibody [E268] ab32019 in HeLa treated with 250ng/ml anisomycin for 30min whole cell lysate.Anti-ATF2 (phospho T71) antibody [E268] ab32019 Immunoprecipitating ATF2 in Human Hela whole cell lysate. For western blotting Anti-ATF2 (phospho T71) antibody [E268] ab32019 (1:1000) was used to confirm successful immunoprecipitation. Blocking and diluting buffer used was 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-ATF2 (phospho T71) antibody [E268] (Anti-ATF2 (phospho T71) antibody [E268] ab32019) at 0.47 µg/mL
Lane 1: HeLa (human cervix adenocarcinoma) treated with 250ng/ml anisomycin for 30min whole cell lysate at 10 µg
Lane 2: HeLa treated with 250ng/ml anisomycin for 30min whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-ATF2 (phospho T71) antibody [E268] ab32019 in HeLa treated with 250ng/ml anisomycin for 30min whole cell lysate
SecondaryAll lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/10000 dilution
Predicted band size: 55 kDa
Exposure time: 3min
This image is courtesy of an anonymous customer review.Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Anti-Brd4 antibody [EPR5150(2)] ab128874 Immunoprecipitating Brd4 in human HEK293 whole cell lysate. 1000μg of cell lysate was incubated with primary antibody (1μg/mg in 50 mM Tris) and matrix (Protein G) for 16 hours at 4°C. For western blotting a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) (1/10000) was used to confirm successful immunoprecipation.
Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366)
BubR1 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with Anti-BubR1 antibody [EPR20652] ab209998 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-BubR1 antibody [EPR20652] ab209998 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1000 dilution.
Lane 1: HeLa whole cell lysate 10 μg (Input).Lane 2: Anti-BubR1 antibody [EPR20652] ab209998 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-BubR1 antibody [EPR20652] ab209998 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.All lanes: Immunoprecipitation - Anti-BubR1 antibody [EPR20652] (Anti-BubR1 antibody [EPR20652] ab209998) at 1/1000 dilution
SecondaryAll lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Predicted band size: 119 kDa
Observed band size: 120 kDa
Exposure time: 1s
Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366)
This data was developed using Anti-SOX9 antibody [EPR12755] ab182579, the same antibody clone in a different buffer formulation.
All lanes: Immunoprecipitation - Anti-SOX9 antibody [EPR12755] (Anti-SOX9 antibody [EPR12755] ab182579) at 1/80 dilution
Lane 1: SW480 (Human colorectal adenocarcinoma epithelial cell) whole cell lysate, 10ug
Lane 2: SW480, 350ug, +, Anti-SOX9 antibody [EPR12755] ab182579 2ug
Lane 3: SW480 cell lysate, 350ug + rabbit IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) , 2ug
SecondaryAll lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 56 kDa
Observed band size: 75 kDa
Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Chk2 was immunoprecipitated from 0.35 mg of NIH/3T3 (mouse embyro fibroblast cell line) whole cell lysate with Anti-Chk2 antibody [EPR19236] ab199031 at 1/40 dilution. Western blot was performed from the immunoprecipitate using Anti-Chk2 antibody [EPR19236] ab199031 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Exposure time 3 minutes.
Lane 1: NIH/3T3 whole cell lysate 10 μg (Input).Lane 2: Anti-Chk2 antibody [EPR19236] ab199031 IP in NIH/3T3 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Chk2 antibody [EPR19236] ab199031 in NIH/3T3 whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-Chk2 antibody [EPR19236] (Anti-Chk2 antibody [EPR19236] ab199031) at 1/1000 dilution
SecondaryAll lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 61 kDa
Observed band size: 61 kDa
Exposure time: 3min
Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366)
GST immunoprecipitation from transfected 293T whole cell lysate using Anti-GST antibody [EPR4236] ab111947. 1 mg of cell lysate was incubated with capture antibody (Anti-GST antibody [EPR4236] ab111947) at 1/20 dilution.
Lane 1: Primary: Anti-GST antibody [EPR4236] ab111947, 1/1000 dilution
Sample: 293T transfected with GST tagged protein vector cell lysate 10 ug
Lane 2: Primary: Anti-GST antibody [EPR4236] ab111947, 1/1000 dilution
Sample: 293T transfected with GST tagged protein vector cell lysate
Lane 3: Primary: Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730, Rabbit monoclonal IgG
Sample: 293T transfected with GST tagged protein vector cell lysateAll lanes: Secondary: ab131366, Veriblot, 1/10000 dilution
Blocking and diluting buffer: 5% NFDM/TBSTLanes 1 - 2: Immunoprecipitation - Anti-GST antibody [EPR4236] (Anti-GST antibody [EPR4236] ab111947) at 1/1000 dilution
Lane 3: Immunoprecipitation - Anti-GST antibody [EPR4236] (Anti-GST antibody [EPR4236] ab111947)
Lane 1: Input - 293 transfected with GST tagged protein vector cell lysate at 10 µg
Lane 2: (+) 293T transfected with GST tagged protein vector cell lysate
Lane 3: (-) 293T transfected with GST tagged protein vector cell lysate
SecondaryAll lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/10000 dilution
Predicted band size: 25 kDa
Observed band size: 41 kDa
This image is courtesy of a customer review submitted by Christian Marx.Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Anti-Bak antibody [Y164] ab32371 immunoprecipitating Bak in human HCT116 p53-/- whole cell lysate. 100μg of cell lysate was incubated with primary antibody (1/100) and matrix (Protein A/G) for 4 hours at 4°C. For western blotting a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) (1/2000) was used to confirm successful immunoprecipation.
Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade ab45166 (purified) at 1/20 immunoprecipitating Histone H4 (acetyl K8) in HeLa treated with Trichostatin A whole cell lysate.
Lane 1 (input): HeLa treated with Trichostatin A whole cell lysate (10μg)
Lane 2 (+): Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade ab45166 + HeLa treated with Trichostatin A whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade ab45166 in HeLa treated with Trichostatin A whole cell lysate.
For western blotting, ab131366 VeriBlot for IP Detection Reagent (HRP) was used for detection (1/10000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Immunoprecipitation - Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade (Anti-Histone H4 (acetyl K8) antibody [EP1002Y] - ChIP Grade ab45166) at 1/20 dilution
Lane 1: HeLa (human cervix adenocarcinoma) treated with Trichostatin A whole cell lysate at 10 µg
Lanes 2 - 3: HeLa (human cervix adenocarcinoma) treated with Trichostatin A whole cell lysate
SecondaryAll lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/10000 dilution
Predicted band size: 11 kDa
Observed band size: 11 kDa
Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBSTAll lanes: Immunoprecipitation - Anti-MX1 antibody [EPR19967] (Anti-MX1 antibody [EPR19967] ab207414) at 1/1000 dilution
Lanes 1 - 2: Daudi (Human Burkitt's lymphoma lymphoblast) treated with 20 U/ml IFN alpha 1 for 24 hours whole cell lysate at 10 µg
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-MX1 antibody [EPR19967] ab207414 in Daudi treated with 20 U/ml IFN alpha 1 for 24 hours whole cell lysate at 10 µg
SecondaryAll lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/5000 dilution
Predicted band size: 76 kDa
Observed band size: 76 kDa
Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366)
All lanes : Anti-Rb (phospho T373) antibody [EP821Y] (Anti-Rb (phospho T373) antibody [EP821Y] ab52975) at 1/500 dilution
Lane 1 : Jurkat (human acute T cell leukemia) starved 24h then 10% FBS incubated for 20h whole cell lysate.
Lane 2 : Jurkat (human acute T cell leukemia ) starved 24h then 10% FBS incubated for 20h whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Rb (phospho T373) antibody [EP821Y] ab52975 in Jurkat (human acute T cell leukemia ) starved 24h then 10% FBS incubated for 20h whole cell lysate.
Lysates/proteins at 10 μg per lane.
Secondary
VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size : 106 kDa
Additional bands at : 110 kDa.
Exposure time : 1 second
Blocking and diluting buffer: 5% NFDM/TBST
The IP lysates are prepared by RIPA method. As the WB image shows, this antibody works in 1%SDS Hot Lysate buffer in WB. So there is no band in input lane.
For Lysate preparation protocol, please refer to the protocol section of the website and/or here.Lanes 1 - 3: Immunoprecipitation - Anti-Rb (phospho T373) antibody [EP821Y] (Anti-Rb (phospho T373) antibody [EP821Y] ab52975) at 1/500 dilution
Lanes 4 - 5: Immunoprecipitation - Anti-Rb (phospho T373) antibody [EP821Y] (Anti-Rb (phospho T373) antibody [EP821Y] ab52975)
Lane 1: Jurkat (human acute T cell leukemia) starved 24h then 10% FBS incubated for 20h whole cell lysate. at 10 µg
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Rb (phospho T373) antibody [EP821Y] ab52975 in Jurkat (human acute T cell leukemia ) starved 24h then 10% FBS incubated for 20h whole cell lysate. at 10 µg
SecondaryAll lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 106 kDa
Observed band size: 110 kDa
Exposure time: 1s
Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366)
MST4 was immunoprecipitated using Anti-MST4 antibody [EP1864Y] ab52491 at 1:100 dilution (2μg in 0.35mg lysates).
All lanes: Immunoprecipitation - Anti-MST4 antibody [EP1864Y] (Anti-MST4 antibody [EP1864Y] ab52491) at 1/1000 dilution
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 10 µg
Lane 2: HeLa whole cell lysate immunoprecipitated with Anti-MST4 antibody [EP1864Y] ab52491 1:100 dilution (2μg in 0.35mg lysates) at 10 µg
Lane 3: HeLa whole cell lysate immunoprecipitated with Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) 1:100 dilution (2μg in 0.35mg lysates), at 10 µg
SecondaryAll lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/5000 dilution
Predicted band size: 47 kDa
Exposure time: 10s
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
This data was developed using Anti-Prothrombin antibody [EPR20159] ab208589, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
Prothrombin is proteolytically cleaved into different fragments. The fragments profile is consistent with what has been described in the literature (PMID: 17637839; 21131592; 16734589).
All lanes: Western blot - Anti-Prothrombin antibody [EPR20159] (Anti-Prothrombin antibody [EPR20159] ab208589) at 1/1000 dilution
Lane 1: Human plasma at 20 µg
Lane 2: Human fetal liver tissue lysate at 20 µg
Lane 3: HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 10 µg
SecondaryAll lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
Predicted band size: 70 kDa
Observed band size: 33 kDa, 52 kDa, 72 kDa
Exposure time: 3min
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Blocking/diluting buffer: 5% NFDM/TBST
This blot was developed using a higher sensitivity ECL substrate.
This antibody detects two forms of Keap1 (α and β) around 74kDa which are consistent with publications PMID: 38397804, PMID: 36142252, PMID: 38242246, etc.
All lanes: Western blot - Anti-Keap1 antibody [EPR22664-26] (Anti-Keap1 antibody [EPR22664-26] ab227828) at 1/1000 dilution
Lane 1: Human lung tissue lysate at 20 µg
Lane 2: Wild-type HAP1 whole cell lysate at 20 µg
Lane 3: Keap1 knockout HAP1 whole cell lysate at 20 µg
Lane 4: HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg
Lane 5: Mouse E14.5 liver tissue lysate at 20 µg
SecondaryLane 1: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Lanes 2 - 5: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 70 kDa
Exposure time: 3min
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-INSL3 antibody [EPR20739] (Anti-INSL3 antibody [EPR20739] ab227974) at 1/10000 dilution
All lanes: Human testis lysate at 10 µg
SecondaryAll lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
Developed using the ECL technique.
Predicted band size: 14 kDa
Observed band size: 15 kDa
Exposure time: 3min
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
This data was developed using Anti-PTGES2/Gbf1 antibody [EPR25169-3] ab300052, the same antibody clone in a different buffer formulation.
Blocking, diluting buffer and concentration was 5% NFDM/TBST.
All lanes: Western blot - Anti-PTGES2/Gbf1 antibody [EPR25169-3] (Anti-PTGES2/Gbf1 antibody [EPR25169-3] ab300052)
Lane 1: Human heart tissue lysate at 20 µg
Lane 2: Human kidney tissue lysate at 20 µg
SecondaryAll lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 42 kDa
Observed band size: 30 kDa
Exposure time: 48s
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
This data was developed using Anti-METTL1 antibody [EPR24320-27] ab271063, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The observed MW is consistent with what has been described in the literature (PMID: 29983320, PMID: 31520064).
Exposure time: 81 seconds
All lanes: Western blot - Anti-METTL1 antibody [EPR24320-27] (Anti-METTL1 antibody [EPR24320-27] ab271063) at 1/1000 dilution
Lane 1: Human kidney tissue lysate at 20 µg
Lane 2: Human pancreas tissue lysate at 20 µg
SecondaryAll lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/2000 dilution
Predicted band size: 31 kDa
Observed band size: 28 kDa, 31 kDa
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
This data was developed using Anti-Decorin antibody [EPR24097-105] ab277636, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The observed MW is consistent with what has been described in the literature (PMID:12601001, 29561836, 8702652, 12623288).
Lower expression tissue: liver (PMID:1611907).
Exposure time: Lane 1: 8 seconds; Lanes 2-4: 48 seconds.
All lanes: Western blot - Anti-Decorin antibody [EPR24097-105] (Anti-Decorin antibody [EPR24097-105] ab277636) at 1/1000 dilution
Lane 1: Human uterus tissue lysate at 20 µg
Lane 2: Human pancreas tissue lysate at 20 µg
Lane 3: Human placenta tissue lysate at 20 µg
Lane 4: Human liver tissue lysate at 20 µg
SecondaryAll lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 40 kDa
Observed band size: 90-140 kDa
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
This data was developed using Anti-160 kD Neurofilament Medium antibody [EPR23510-76] ab254348, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The molecular weight is consistent to what has been described in the literature (PMID:19239416, PMID:27000625).
Negative control: liver (PMID:30541916).
Exposure time: 10 secs.
All lanes: Western blot - Anti-160 kD Neurofilament Medium antibody [EPR23510-76] (Anti-160 kD Neurofilament Medium antibody [EPR23510-76] ab254348) at 1/1000 dilution
Lane 1: Human cerebellum tissue lysate at 20 µg
Lane 2: Human nerve tissue lysate at 20 µg
Lane 3: Human liver tissue lysate at 20 µg
SecondaryAll lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 102 kDa
Observed band size: 160 kDa
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
This data was developed using Anti-Nicotinic Acetylcholine Receptor alpha 5/CHRNA5 antibody [EPR24135-98] ab259859, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The observed MW is consistent with what has been described in the literature (PMID:30543688).
Low expression: kidney, liver (PMID:20920278).
Exposure time: Lane 1: 10 seconds
Lane 2-5: 26 seconds
All lanes: Western blot - Anti-Nicotinic Acetylcholine Receptor alpha 5/CHRNA5 antibody [EPR24135-98] (Anti-Nicotinic Acetylcholine Receptor alpha 5/CHRNA5 antibody [EPR24135-98] ab259859) at 1/1000 dilution
Lane 1: Human Hippocampus tissue lysate at 20 µg
Lane 2: Human brain tissue lysate at 20 µg
Lane 3: Human testis tissue lysate at 20 µg
Lane 4: Human kidney tissue lysate at 20 µg
Lane 5: Human liver tissue lysate at 20 µg
SecondaryAll lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 53 kDa
Observed band size: 53 kDa
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Low expression: human ovary (PMID:2328255).
The bands nearby 25 kDa could be non-specific bands.
Exposure time: Lane 1-3: 3.25 seconds Lane 4-5: 37 seconds
All lanes: Western blot - Anti-Thymidine Phosphorylase antibody [EPR25629-129] (Anti-Thymidine Phosphorylase antibody [EPR25629-129] ab284861) at 1/1000 dilution
Lane 1: Human tonsil tissue lysate at 20 µg
Lane 2: human ovary cancer tissue lysate at 20 µg
Lane 3: human ovary tissue lysate at 20 µg
Lane 4: human colon tissue lysate at 20 µg
Lane 5: human liver tissue lysate at 20 µg
SecondaryLanes 1 - 5: VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution
Lanes 1 - 5: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 50 kDa
Observed band size: 50 kDa
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
This data was developed using Anti-ASAH1 antibody [EPR24476-30] ab282276, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID:7744740, 26553872, 30254208).
Negative control: A375 (PMID: 26553872, 30254208); MDA-MB-231 (PMID: 26553872).
Exposure time: 37 seconds
All lanes: Western blot - Anti-ASAH1 antibody [EPR24476-30] (Anti-ASAH1 antibody [EPR24476-30] ab282276) at 1/1000 dilution
Lane 1: SK-MEL-28 (human malignant melanoma) whole cell lysate at 20 µg
Lane 2: MeWo (human malignant melanoma fibroblast) whole cell lysate at 20 µg
Lane 3: A375 (human malignant melanoma epithelial cell) whole cell lysate at 20 µg
Lane 4: MDA-MB-231 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg
SecondaryAll lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/5000 dilution
Predicted band size: 45 kDa
Observed band size: 37 kDa
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Blocking/Dilution buffer: 5% NFDM/TBST.
Negative control: Human liver (PMID:20191564).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-beta III Tubulin antibody [AA10] ab231083).
All lanes: Western blot - Anti-beta III Tubulin antibody [AA10] (Anti-beta III Tubulin antibody [AA10] ab231083) at 1/1000 dilution
Lane 1: Human brain tissue lysate at 10 µg
Lane 2: Human heart tissue lysate at 10 µg
SecondaryAll lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 50 kDa
Exposure time: 3s
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
This data was developed using 259980, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Negative control: mouse embryo/fetal liver (PIMID:16894625).
Exposure time: 70 seconds
All lanes: Western blot - Anti-TBX5 antibody [EPR23798-102] (Anti-TBX5 antibody [EPR23798-102] ab259980) at 1/1000 dilution
Lane 1: Mouse P0 heart tissue lysate at 20 µg
Lane 2: Mouse P0 liver tissue lysate at 20 µg
Lane 3: Mouse E18 heart tissue lysate at 20 µg
Lane 4: Mouse E18 liver tissue lysate at 20 µg
Lane 5: Mouse E17 heart tissue lysate at 20 µg
Lane 6: Mouse E17 liver tissue lysate at 20 µg
SecondaryAll lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 58 kDa
Observed band size: 57 kDa
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Blocking and dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with what has been described in the literature (PMID:26654173, 30123430).
All lanes: Western blot - Anti-Orai3 antibody [EPR22575-17] (Anti-Orai3 antibody [EPR22575-17] ab254260) at 1/1000 dilution
Lane 1: Human kidney lysate at 20 µg
Lane 2: Human testis lysate at 20 µg
Lane 3: MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 4: A549 (human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 5: RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate at 10 µg
Lane 6: NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 10 µg
SecondaryLanes 1 - 2: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Lanes 3 - 6: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 31 kDa
Observed band size: 33 kDa
Exposure time: 3min
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
This data was developed using Anti-Matrin 3 antibody [Z-MATR3-4] - Mouse IgG2a (Chimeric) ab281927, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 26878116).
This blot was developed using a higher sensitivity ECL substrate.
Exposure time: 84 seconds.
All lanes: Western blot - Anti-Matrin 3 antibody [Z-MATR3-4] - Mouse IgG2a (Anti-Matrin 3 antibody [Z-MATR3-4] - Mouse IgG2a (Chimeric) ab281927) at 1/1000 dilution
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 10 µg
Lane 2: K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate at 10 µg
SecondaryAll lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 95 kDa
Observed band size: 120 kDa
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Blocking and dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-non-muscle Myosin IIA antibody [EPR22933-9] (Anti-non-muscle Myosin IIA antibody [EPR22933-9] ab238131) at 1/1000 dilution
Lane 1: Human liver tissue lysate 20ug
Lane 2: Human lung lysate at 20 µg
Lane 3: Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate at 20 µg
Lane 4: HEK-293T (human embryonic kidney epithelial cell), whole cell lysate at 20 µg
Lane 5: HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg
Lane 6: Mouse lung tissue lysate at 20 µg
Lane 7: Rat spleen tissue lysate at 20 µg
Lane 8: Rat lung tissue lysate at 20 µg
SecondaryLanes 1 - 3: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Lanes 4 - 8: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 227 kDa
Observed band size: 226 kDa
Exposure time: 26s
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
This data was developed using Anti-Granzyme K antibody [EPR24601-164] ab282703, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The expression profile/molecular weight observed is consistent with what has been described in the literature (PMID: 10407175, 18559365).
Negative control: heart (PMID: 10407175).
Exposure time: 15 seconds
All lanes: Western blot - Anti-Granzyme K antibody [EPR24601-164] (Anti-Granzyme K antibody [EPR24601-164] ab282703) at 1/1000 dilution
Lane 1: Human spleen tissue lysate at 40 µg
Lane 2: Human tonsil tissue lysate at 40 µg
Lane 3: Human lymphoma tissue lysate at 40 µg
Lane 4: Human heart tissue lysate at 40 µg
SecondaryAll lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/2000 dilution
Predicted band size: 29 kDa
Observed band size: 22 kDa, 27 kDa
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
This data was developed using 282577, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The expression pattern & observed MW are consistent with what has been described in the literature (PMID: 8999038).
Low expression: human tonsil (PMID: 8999038).
Exposure time: 3 minutes
All lanes: Western blot - Anti-OSMR antibody [EPR24611-71] (Anti-OSMR antibody [EPR24611-71] ab282577) at 1/1000 dilution
Lane 1: Human placenta tissue lysate at 20 µg
Lane 2: Human tonsil tissue lysate at 20 µg
SecondaryAll lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/2000 dilution
Predicted band size: 111 kDa
Observed band size: 150-180 kDa
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Blocking/Dilution buffer: 5% NFDM/TBST.
This blot was developed using a higher sentivity ECL substrate.
The molecular weight observed is consistent with what has been described in the literature (PMID:30118527; PMID:1662607).
All lanes: Western blot - Anti-15 Lipoxygenase 1 antibody [EPR22136] (Anti-15 Lipoxygenase 1 antibody [EPR22136] ab242062) at 1/1000 dilution
All lanes: Human colon at 20 µg
SecondaryAll lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 75 kDa
Observed band size: 74 kDa
Exposure time: 114s
Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366)
Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
Exposure Time: Lane 1: 7 secs; Lane 2: 3 mins; Lane 3: 3 secs; Lane 4: 3 mins.
The molecular weight observed is consistent with literatures (PMID: PMID:20070425, PMID:18256920, PMID:18938162).
All lanes: Western blot - Anti-CD200 / OX2 antibody [EPR22412-229] (Anti-CD200 / OX2 antibody [EPR22412-229] ab254193) at 1/1000 dilution
Lane 1: Human brain tissue lysate at 20 µg
Lane 2: Human lung tissue lysate at 20 µg
Lane 3: Human placenta tissue lysate at 20 µg
Lane 4: Human tonsil tissue lysate at 20 µg
SecondaryAll lanes: Western blot - VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/100000 dilution
Predicted band size: 31 kDa
Observed band size: 40-50 kDa
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