Log In Sign up
The largest database of trusted experimental protocols

Lamelli buffer

Manufactured by Bio-Rad
Visit Supplier

Lamelli buffer is a protein sample preparation buffer used in gel electrophoresis. It denatures proteins and ensures their even distribution within the gel matrix.

Automatically generated - may contain errors

Lab products found in correlation

Protein g dynabead, by Thermo Fisher Scientific (3 mentions) Whole mouse igg, by Merck Group (3 mentions) Flag m2, by Merck Group (3 mentions) Ip lysis buffer, by Bio-Rad (2 mentions) Proteinase and phosphatase inhibitors, by Merck Group (1 mentions) β mercaptoethanol, by Merck Group (1 mentions) Ripa buffer, by Thermo Fisher Scientific (1 mentions) Native sample buffer, by Bio-Rad (1 mentions) Bovine serum albumin lyophilized powder, by Merck Group (1 mentions) Kanamycin, by Merck Group (1 mentions) N lauroylsarcosine sodium salt, by Merck Group (1 mentions) N decyl β d maltopyranoside, by Anatrace (1 mentions) O nitrobenzaldehyde, by Merck Group (1 mentions) Mini protean precast gel, by Bio-Rad (1 mentions) Amicon ultra 0.5 centrifugal filter unit, by Merck Group (1 mentions) Ampicillin, by Merck Group (1 mentions) Triton x 100, by Merck Group (1 mentions) Lb broth, by Merck Group (1 mentions) Protease inhibitor, by Thermo Fisher Scientific (1 mentions) Phosphatase inhibitor, by Merck Group (1 mentions)

Close spelling variants of this product

4x Lamelli sample buffer

7 protocols using lamelli buffer

1

GATA6 Protein Co-Immunoprecipitation

Check if the same lab product or an alternative is used in the 5 most similar protocols
GATA6Ex4Δ1/Δ2:ind GATA6 cells were differentiated to mesendoderm (day 2). Between days 1 and 2, the cells were cultured in the absence or presence of 10ng/ml doxycycline to induce GATA6–3xFLAG expression at endogenous levels. At Day 2, cells were washed once with PBS and lysed with IP lysis buffer (ThermoFisher/Pierce, IL, #87787) with HALT proteinase inhibitor cocktail (ThermoFisher/Pierce, IL, #78443) and Universal Nuclease (ThermoFisher/Pierce, IL, #88701). The sample was incubated at room temperature for 10 minutes before centrifugation to remove cell debris. The lysate was pre-cleared with pre-washed Protein G Dynabeads (ThermoFisher Scientific, NY, #10004D) for 2 hours at 4°C. An input sample was taken and the remaining lysate split equally and incubated with either 1 μg of FLAG M2 (Sigma-Aldrich, MO, #F1804) or 1 μg whole mouse IgG (Sigma-Aldrich, MO, #12–371) antibodies overnight at 4°C. The following day, pre-washed Protein G Dynabeads were added and the mixture was incubated for 2 hours at 4°C. The beads were then washed 5 times with IP lysis buffer and eluted in 1x Lamelli buffer (BioRad, CA, #1610747) at 95°C for 10 minutes. Samples were then analyzed by western blot. Antibodies used for co-IP listed in Table S8.
Heslop J.A., Pournasr B., Liu J.T, & Duncan S.A. (2021). GATA6 defines endoderm fate by controlling chromatin accessibility during differentiation of human-induced pluripotent stem cells. Cell reports, 35(7), 109145.
+ Open protocol
+ Expand
2

Protein Detection by Western Blotting

Check if the same lab product or an alternative is used in the 5 most similar protocols
For the detection of proteins, cells were lysed with 50–100 µl of RIPA buffer (Pierce Biotechnology), supplemented with proteinase and phosphatase inhibitors (EMD Millipore). Samples were incubated on ice for 30 min and were intermittently vortexed and sonicated (three bursts of 10 s at 3 W). Lysates were cleared by centrifugation in a tabletop centrifuge (16,000 × g, 10 min, 4 °C). Supernatants were transferred to fresh tubes and protein concentration was measured by means of Bradford protein assays.
Equal amounts of proteins were mixed with Lamelli Buffer (Bio-Rad) supplemented with 5% beta-mercaptoethanol (Sigma-Aldrich). Samples were denatured by incubation at 99 °C for 10 min. Samples were then loaded on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gels and subsequently weight-separated by electrophoresis. Separated proteins were then transferred to Nitrocellulose membranes with the Fast-Transfer apparatus and kit (Bio-Rad). All antibodies used in this study are listed in Supplementary Table 4. Unless otherwise stated, primary antibodies were used in a 1:1000 and secondary antibodies in a 1:5000 dilution.
Papaioannou D., Petri A., Dovey O.M., Terreri S., Wang E., Collins F.A., Woodward L.A., Walker A.E., Nicolet D., Pepe F., Kumchala P., Bill M., Walker C.J., Karunasiri M., Mrózek K., Gardner M.L., Camilotto V., Zitzer N., Cooper J.L., Cai X., Rong-Mullins X., Kohlschmidt J., Archer K.J., Freitas M.A., Zheng Y., Lee R.J., Aifantis I., Vassiliou G., Singh G., Kauppinen S., Bloomfield C.D., Dorrance A.M, & Garzon R. (2019). The long non-coding RNA HOXB-AS3 regulates ribosomal RNA transcription in NPM1-mutated acute myeloid leukemia. Nature Communications, 10, 5351.
+ Open protocol
+ Expand
3

Purification and Analysis of CR8020 Antibody

Check if the same lab product or an alternative is used in the 5 most similar protocols
Mini-PROTEAN precast gels, Lamelli buffer, native sample buffer, and HRP enzyme substrate are obtained from Bio-Rad Laboratories. BL21(DE3), Rosetta (DE3) PLysS, and Color Prestained Protein Standard are obtained from New England Bio-Laboratories. Ultracel-3 regenerated cellulose membranes and Amicon Ultra-0.5 centrifugal Filter Units are from Millipore Sigma. o-Nitrobenzaldehyde, bovine serum albumin lyophilized powder, ≥96% (agarose gel electrophoresis), LB broth, ampicillin, kanamycin, N-lauroylsarcosine sodium salt, and Triton X-100 are from Sigma-Aldrich. n-Decyl β-d-maltopyranoside (DM) is from Anatrace. All lipids are from Avanti Polar Lipids. Goat anti-human IgG (H+L) HRP is purchased from Invitrogen. Nonfat dry milk is obtained from Lab Scientific. CR8020 is a generous gift from M. Crank at the National Institute of Allergy and Infectious Diseases (NIAID).
Eller M.W., Siaw H.M, & Dyer R.B. (2021). Stability of HA2 Prefusion Structure and pH-Induced Conformational Changes in the HA2 Domain of H3N2 Hemagglutinin. Biochemistry, 60(35), 2623-2636.
+ Open protocol
+ Expand
4

Bacterial Protein Extraction and Denaturation

Check if the same lab product or an alternative is used in the 5 most similar protocols
One milliliter of bacteria for each corresponding supernatant was centrifuged at 14,000 rpm for 15 min at 4°C to pellet the bacteria. Supernatant was removed and bacteria were resuspended in 1:1 ratio of dH2O and 2× Lamelli buffer (BioRad) containing 10% β-mercaptoethanol. Samples were then heated at 95°C for 10 min for protein denaturation.
Gabor C.E., Hazen T.H., Delaine-Elias B.C., Rasko D.A, & Barry E.M. (2023). Genomic, transcriptomic, and phenotypic differences among archetype Shigella flexneri strains of serotypes 2a, 3a, and 6. mSphere, 8(6), e00408-23.
+ Open protocol
+ Expand
5

Quantitative Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
For western blots, protein was collected using RIPA buffer (Life Technologies 89900), protease inhibitor (Thermo 1860932) and phosphatase inhibitor (Sigma 04906837001) and quantified using Qubit protein assay kit (Thermo Q33211). SDS-PAGE was run using 25–50 μg of protein mixed 1:1 with lamelli buffer (BioRad 161–0737) and heated to 90°C for 10 minutes and loaded onto a Mini-PROTEAN TGX gel (BioRad 456–8094). After gel electrophoresis, samples were transferred to TransBlot Turbo Transfer Pack 0.2 μm PVDF membrane (BioRad 1704156) using TransBlot Turbo (BioRad). Membrane was blocked using 5% BSA blocking solution for 30 minutes then primary antibodies were added at a 1:100 concentration overnight at 4C. The primary antibodies used were as follows: Anti-TEK (RD AF762), Anti-ANGPT2 (RD AF623), Anti-SMAD4 (Abcam ab40759), Anti-βACTIN (Cell Signaling 3700). After washing, appropriate LICOR IRDye 800CW or 680RD secondary antibodies were used at 1:1000 and incubated at room temperature for 1 hour. Membranes were imaged using Odyessy (LICOR) and iStudio Lite was used for western quantification.
Crist A.M., Zhou X., Garai J., Lee A.R., Thoele J., Ullmer C., Klein C., Zabaleta J, & Meadows S.M. (2019). Angiopoietin-2 Inhibition Rescues Arteriovenous Malformation in a Smad4 Hereditary Hemorrhagic Telangiectasia Mouse Model. Circulation, 139(17), 2049-2063.
+ Open protocol
+ Expand
6

GATA6 Knockout Differentiation Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
GATA6-/- cells were differentiated to day 2 of the Duncan lab differentiation protocol. After 24 h of differentiation, the cells were cultured in the absence or presence of doxycycline. Samples were collected at day 2. Cells were lysed with IP lysis buffer (ThermoFisher/Pierce, IL, #87787) containing 1x HALT proteinase inhibitor cocktail (ThermoFisher/Pierce, IL, #78443) and Universal Nuclease (ThermoFisher/Pierce, IL, #88701). Lysates were pre-cleared with Protein G Dynabeads (ThermoFisher Scientific, NY, #10004D) for 2 h at 4°C with shaking. The precleared samples were split equally and incubated with either 1 μg of FLAG M2 (Sigma-Aldrich, MO, #F1804) or 1 μg whole mouse IgG (Sigma-Aldrich, MO, #12-371) antibodies overnight at 4°C. Protein G Dynabeads were then added and the mixture was incubated for 2 h at 4°C. IP lysis buffer was used to wash the beads five times, before elution in 1x Lamelli buffer (BioRad, CA, #1610747) at 95°C for 10 min before analysis by western blot.
Heslop J.A., Pournasr B, & Duncan S.A. (2022). Chromatin remodeling is restricted by transient GATA6 binding during iPSC differentiation to definitive endoderm. iScience, 25(5), 104300.
+ Open protocol
+ Expand
7

GATA6 Protein Co-Immunoprecipitation

Check if the same lab product or an alternative is used in the 5 most similar protocols
GATA6Ex4Δ1/Δ2:ind GATA6 cells were differentiated to mesendoderm (day 2). Between days 1 and 2, the cells were cultured in the absence or presence of 10ng/ml doxycycline to induce GATA6–3xFLAG expression at endogenous levels. At Day 2, cells were washed once with PBS and lysed with IP lysis buffer (ThermoFisher/Pierce, IL, #87787) with HALT proteinase inhibitor cocktail (ThermoFisher/Pierce, IL, #78443) and Universal Nuclease (ThermoFisher/Pierce, IL, #88701). The sample was incubated at room temperature for 10 minutes before centrifugation to remove cell debris. The lysate was pre-cleared with pre-washed Protein G Dynabeads (ThermoFisher Scientific, NY, #10004D) for 2 hours at 4°C. An input sample was taken and the remaining lysate split equally and incubated with either 1 μg of FLAG M2 (Sigma-Aldrich, MO, #F1804) or 1 μg whole mouse IgG (Sigma-Aldrich, MO, #12–371) antibodies overnight at 4°C. The following day, pre-washed Protein G Dynabeads were added and the mixture was incubated for 2 hours at 4°C. The beads were then washed 5 times with IP lysis buffer and eluted in 1x Lamelli buffer (BioRad, CA, #1610747) at 95°C for 10 minutes. Samples were then analyzed by western blot. Antibodies used for co-IP listed in Table S8.
Heslop J.A., Pournasr B., Liu J.T, & Duncan S.A. (2021). GATA6 defines endoderm fate by controlling chromatin accessibility during differentiation of human-induced pluripotent stem cells. Cell reports, 35(7), 109145.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!