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4x laemmli sample buffer bio rad  (Bio-Rad)


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    Bio-Rad 4x laemmli sample buffer bio rad
    Targeted activation of endogenous mitochondrial ClpP. (A) Western blots of mitochondrial lysates (30 μg) from wild‐type and ClpP‐KO HEK293T cells were probed with antibodies against ClpP and α‐tubulin, the latter being used as a loading control. (B) Analysis of ClpP and ClpX assembly was carried out in wild‐type HEK293T mitochondria through isokinetic sucrose gradient centrifugation. Mitochondrial lysate (0.65 mg) was centrifuged on a 10%–30% gradient using the SW 60Ti rotor at 40 000 rpm for 2 h and 15 min at 4°C. Small and large mitoribosomal subunits were used as a reference of migration and were immunodecorated by antibodies against marker proteins of the mt‐SSU (mS29; HUGO protein name: DAP3) and the mt‐LSU (uL13m; HUGO protein name: MRPL13). (C) Effect of ONC201‐mediated ClpP activation on casein hydrolysis in the PAGE‐based fluorescent assay. Mitochondrial lysates (50 μg) from wild‐type and KO cells were used as a source of ClpP activity, and incubated with ONC201 at a final concentration of 100 μM. Upper panels: Milk casein isoforms (α, β and k) are depicted to the left; molecular weight markers (M; Precision Plus Protein Dual Color Standards, <t>Bio‐Rad)</t> are indicated to the right. Middle panels: Total densitometric profile of fluorescent substrate was determined for each lane. Lower panels: Coomassie blue staining (CBS) of mitochondrial proteins was used as a loading control. (D) Chemical structure of DA29 (ClpP activator) and DA28 (negative control). (E) Effect of ClpP chemoactivation by DA29 on casein hydrolysis in the PAGE‐based fluorescent assay; DA28 was used as a negative control. For description, see panel C.
    4x Laemmli Sample Buffer Bio Rad, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 11850 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/4x+laemmli+sample+buffer/4x+Laemmli+Sample+Buffer/pmc13675934-66-17-21
    Average 99 stars, based on 11850 article reviews
    4x laemmli sample buffer bio rad - by Bioz Stars, 2026-10
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    1) Product Images from "Elucidating the Molecular Impact of DA29‐Induced ClpP Chemoactivation on Mitochondrial Function"

    Article Title: Elucidating the Molecular Impact of DA29‐Induced ClpP Chemoactivation on Mitochondrial Function

    Journal: The FASEB Journal

    doi: 10.1096/fj.202603398RR

    Targeted activation of endogenous mitochondrial ClpP. (A) Western blots of mitochondrial lysates (30 μg) from wild‐type and ClpP‐KO HEK293T cells were probed with antibodies against ClpP and α‐tubulin, the latter being used as a loading control. (B) Analysis of ClpP and ClpX assembly was carried out in wild‐type HEK293T mitochondria through isokinetic sucrose gradient centrifugation. Mitochondrial lysate (0.65 mg) was centrifuged on a 10%–30% gradient using the SW 60Ti rotor at 40 000 rpm for 2 h and 15 min at 4°C. Small and large mitoribosomal subunits were used as a reference of migration and were immunodecorated by antibodies against marker proteins of the mt‐SSU (mS29; HUGO protein name: DAP3) and the mt‐LSU (uL13m; HUGO protein name: MRPL13). (C) Effect of ONC201‐mediated ClpP activation on casein hydrolysis in the PAGE‐based fluorescent assay. Mitochondrial lysates (50 μg) from wild‐type and KO cells were used as a source of ClpP activity, and incubated with ONC201 at a final concentration of 100 μM. Upper panels: Milk casein isoforms (α, β and k) are depicted to the left; molecular weight markers (M; Precision Plus Protein Dual Color Standards, Bio‐Rad) are indicated to the right. Middle panels: Total densitometric profile of fluorescent substrate was determined for each lane. Lower panels: Coomassie blue staining (CBS) of mitochondrial proteins was used as a loading control. (D) Chemical structure of DA29 (ClpP activator) and DA28 (negative control). (E) Effect of ClpP chemoactivation by DA29 on casein hydrolysis in the PAGE‐based fluorescent assay; DA28 was used as a negative control. For description, see panel C.
    Figure Legend Snippet: Targeted activation of endogenous mitochondrial ClpP. (A) Western blots of mitochondrial lysates (30 μg) from wild‐type and ClpP‐KO HEK293T cells were probed with antibodies against ClpP and α‐tubulin, the latter being used as a loading control. (B) Analysis of ClpP and ClpX assembly was carried out in wild‐type HEK293T mitochondria through isokinetic sucrose gradient centrifugation. Mitochondrial lysate (0.65 mg) was centrifuged on a 10%–30% gradient using the SW 60Ti rotor at 40 000 rpm for 2 h and 15 min at 4°C. Small and large mitoribosomal subunits were used as a reference of migration and were immunodecorated by antibodies against marker proteins of the mt‐SSU (mS29; HUGO protein name: DAP3) and the mt‐LSU (uL13m; HUGO protein name: MRPL13). (C) Effect of ONC201‐mediated ClpP activation on casein hydrolysis in the PAGE‐based fluorescent assay. Mitochondrial lysates (50 μg) from wild‐type and KO cells were used as a source of ClpP activity, and incubated with ONC201 at a final concentration of 100 μM. Upper panels: Milk casein isoforms (α, β and k) are depicted to the left; molecular weight markers (M; Precision Plus Protein Dual Color Standards, Bio‐Rad) are indicated to the right. Middle panels: Total densitometric profile of fluorescent substrate was determined for each lane. Lower panels: Coomassie blue staining (CBS) of mitochondrial proteins was used as a loading control. (D) Chemical structure of DA29 (ClpP activator) and DA28 (negative control). (E) Effect of ClpP chemoactivation by DA29 on casein hydrolysis in the PAGE‐based fluorescent assay; DA28 was used as a negative control. For description, see panel C.

    Techniques Used: Activation Assay, Western Blot, Control, Analysis, Gradient Centrifugation, Migration, Marker, Fluorescence, Activity Assay, Incubation, Concentration Assay, Molecular Weight, Staining, Negative Control

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    Article Snippet: Total protein concentration was quantified by BCA assay (Thermo Scientific). .. Thirty micrograms of protein extracts were diluted in 4X laemmli sample buffer (Bio-Rad) containing DTT (Sigma) and denatured at 95 °C for 5 min. Proteins were separated on 4-12% gradient SDS-PAGE gels (BioRad) and transferred to nitrocellulose membranes (Bio-Rad). .. The blots were stained with Ponceau S Staining solution (Cell Signaling) and following blocking with 5% BSA in PBST for 1 h at RT, membranes were incubated with primary antibodies in 5% BSA in PBST overnight at 4°C with agitation.

    Article Title: Small extracellular vesicles as mediators of paracrine senescence in cellular models of aging.
    Article Snippet: sEV were lysed using the following lysis buffer [(TrisHCl 20 mM pH 7.6; DTT 1 mM; EDTA 1 mM; PMSF 1 mM; benzamidine 1 mM; sodium molybdate 2 mM; β-sodium glycerophosphate 2 mM; sodium orthovanadate 0.2 mM; KCl 120 mM; 1 mg/ml (each) leupeptin, pepstatin A and antipain; Nonidet P-40 0.5% (v/v); Triton X-100 0.1% (v/v)] (all from Sigma- Aldrich)]. .. Lysates were diluted with 4X Laemmli Sample Buffer (Bio-Rad, UK) and an equal number of particles (3x109) were separated in SDS-PAGE gels at 80V. ..

    other:

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    Centrifugation:

    Article Title: Mechanisms of transcription control by distal enhancers from single-gene nanoscopy.
    Article Snippet: mESCs were lysed in RIPA buffer (50 mM Tris–HCl pH 7.4, 150 mM NaCl, 1% Triton X-100, 0.5% sodium deoxycholate, and 0.1% SDS) supplemented with protease inhibitors (Thermo fisher, A32965) for 30 min at 4 ◦C. .. Lysates were clarified by centrifugation at 12,000 g for 5 min, and the supernatants were mixed with 4x Laemmli Sample Buffer (Bio-Rad, 1610747) and boiled at 95 ◦C for 5 min. Proteins were resolved on 4–20% SDS–PAGE gels (Bio-Rad) at 120 V for 1 hr and transferred to 0.2 μm PVDF membranes (Bio-Rad,1620177) at 300 mA for 80 min. Membranes were blocked with 3% non-fat milk (CST, 9999) in TBST for 1 hr at room temperature and incubated with primary antibodies overnight at 4 ◦C. ..

    Incubation:

    Article Title: Mechanisms of transcription control by distal enhancers from single-gene nanoscopy.
    Article Snippet: mESCs were lysed in RIPA buffer (50 mM Tris–HCl pH 7.4, 150 mM NaCl, 1% Triton X-100, 0.5% sodium deoxycholate, and 0.1% SDS) supplemented with protease inhibitors (Thermo fisher, A32965) for 30 min at 4 ◦C. .. Lysates were clarified by centrifugation at 12,000 g for 5 min, and the supernatants were mixed with 4x Laemmli Sample Buffer (Bio-Rad, 1610747) and boiled at 95 ◦C for 5 min. Proteins were resolved on 4–20% SDS–PAGE gels (Bio-Rad) at 120 V for 1 hr and transferred to 0.2 μm PVDF membranes (Bio-Rad,1620177) at 300 mA for 80 min. Membranes were blocked with 3% non-fat milk (CST, 9999) in TBST for 1 hr at room temperature and incubated with primary antibodies overnight at 4 ◦C. ..

    Western Blot:

    Article Title: Lycopene inhibits proliferation, induces apoptosis, and suppresses migration in cervical cancer cells: involvement of PI3K/Akt signaling.
    Article Snippet: 1 Laboratorio de Ciencias de la Naturaleza, Departamento de Ciencias de la Naturaleza, Centro Universitario del Sur (CUSur), Universidad de Guadalajara, Ciudad Guzmán, Jalisco, México 2 Departamento de Fisiología, Instituto Nacional de Cardiología Ignacio Chávez, Juan Badiano 1, Sección XVI, Tlalpan, 14080 Ciudad de Mexico, México 3 Centro de Investigación, Facultad de Medicina, Facultad de Medicina y Cirugía, UNAM-UABJO, Universidad Autónoma “Benito Juárez” de Oaxaca, 68020 Ciudad Oaxaca de Juárez, México Abstract Cervical cancer remains a major cause of mortality worldwide, highlighting the need for alternative therapeutic strategies.. Lycopene, a dietary carotenoid with known antioxidant properties, has demonstrated anticancer activity in several malignancies; however, its mechanistic effects in cervical cancer remain poorly understood.. We hypothesize that lycopene inhibits the growth of cervical cancer cell lines and certain cancer-related processes.

    Electrophoresis:

    Article Title: Lycopene inhibits proliferation, induces apoptosis, and suppresses migration in cervical cancer cells: involvement of PI3K/Akt signaling.
    Article Snippet: 1 Laboratorio de Ciencias de la Naturaleza, Departamento de Ciencias de la Naturaleza, Centro Universitario del Sur (CUSur), Universidad de Guadalajara, Ciudad Guzmán, Jalisco, México 2 Departamento de Fisiología, Instituto Nacional de Cardiología Ignacio Chávez, Juan Badiano 1, Sección XVI, Tlalpan, 14080 Ciudad de Mexico, México 3 Centro de Investigación, Facultad de Medicina, Facultad de Medicina y Cirugía, UNAM-UABJO, Universidad Autónoma “Benito Juárez” de Oaxaca, 68020 Ciudad Oaxaca de Juárez, México Abstract Cervical cancer remains a major cause of mortality worldwide, highlighting the need for alternative therapeutic strategies.. Lycopene, a dietary carotenoid with known antioxidant properties, has demonstrated anticancer activity in several malignancies; however, its mechanistic effects in cervical cancer remain poorly understood.. We hypothesize that lycopene inhibits the growth of cervical cancer cell lines and certain cancer-related processes.

    Blocking Assay:

    Article Title: Lycopene inhibits proliferation, induces apoptosis, and suppresses migration in cervical cancer cells: involvement of PI3K/Akt signaling.
    Article Snippet: 1 Laboratorio de Ciencias de la Naturaleza, Departamento de Ciencias de la Naturaleza, Centro Universitario del Sur (CUSur), Universidad de Guadalajara, Ciudad Guzmán, Jalisco, México 2 Departamento de Fisiología, Instituto Nacional de Cardiología Ignacio Chávez, Juan Badiano 1, Sección XVI, Tlalpan, 14080 Ciudad de Mexico, México 3 Centro de Investigación, Facultad de Medicina, Facultad de Medicina y Cirugía, UNAM-UABJO, Universidad Autónoma “Benito Juárez” de Oaxaca, 68020 Ciudad Oaxaca de Juárez, México Abstract Cervical cancer remains a major cause of mortality worldwide, highlighting the need for alternative therapeutic strategies.. Lycopene, a dietary carotenoid with known antioxidant properties, has demonstrated anticancer activity in several malignancies; however, its mechanistic effects in cervical cancer remain poorly understood.. We hypothesize that lycopene inhibits the growth of cervical cancer cell lines and certain cancer-related processes.



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    Targeted activation of endogenous mitochondrial ClpP. (A) Western blots of mitochondrial lysates (30 μg) from wild‐type and ClpP‐KO HEK293T cells were probed with antibodies against ClpP and α‐tubulin, the latter being used as a loading control. (B) Analysis of ClpP and ClpX assembly was carried out in wild‐type HEK293T mitochondria through isokinetic sucrose gradient centrifugation. Mitochondrial lysate (0.65 mg) was centrifuged on a 10%–30% gradient using the SW 60Ti rotor at 40 000 rpm for 2 h and 15 min at 4°C. Small and large mitoribosomal subunits were used as a reference of migration and were immunodecorated by antibodies against marker proteins of the mt‐SSU (mS29; HUGO protein name: DAP3) and the mt‐LSU (uL13m; HUGO protein name: MRPL13). (C) Effect of ONC201‐mediated ClpP activation on casein hydrolysis in the PAGE‐based fluorescent assay. Mitochondrial lysates (50 μg) from wild‐type and KO cells were used as a source of ClpP activity, and incubated with ONC201 at a final concentration of 100 μM. Upper panels: Milk casein isoforms (α, β and k) are depicted to the left; molecular weight markers (M; Precision Plus Protein Dual Color Standards, <t>Bio‐Rad)</t> are indicated to the right. Middle panels: Total densitometric profile of fluorescent substrate was determined for each lane. Lower panels: Coomassie blue staining (CBS) of mitochondrial proteins was used as a loading control. (D) Chemical structure of DA29 (ClpP activator) and DA28 (negative control). (E) Effect of ClpP chemoactivation by DA29 on casein hydrolysis in the PAGE‐based fluorescent assay; DA28 was used as a negative control. For description, see panel C.
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    Targeted activation of endogenous mitochondrial ClpP. (A) Western blots of mitochondrial lysates (30 μg) from wild‐type and ClpP‐KO HEK293T cells were probed with antibodies against ClpP and α‐tubulin, the latter being used as a loading control. (B) Analysis of ClpP and ClpX assembly was carried out in wild‐type HEK293T mitochondria through isokinetic sucrose gradient centrifugation. Mitochondrial lysate (0.65 mg) was centrifuged on a 10%–30% gradient using the SW 60Ti rotor at 40 000 rpm for 2 h and 15 min at 4°C. Small and large mitoribosomal subunits were used as a reference of migration and were immunodecorated by antibodies against marker proteins of the mt‐SSU (mS29; HUGO protein name: DAP3) and the mt‐LSU (uL13m; HUGO protein name: MRPL13). (C) Effect of ONC201‐mediated ClpP activation on casein hydrolysis in the PAGE‐based fluorescent assay. Mitochondrial lysates (50 μg) from wild‐type and KO cells were used as a source of ClpP activity, and incubated with ONC201 at a final concentration of 100 μM. Upper panels: Milk casein isoforms (α, β and k) are depicted to the left; molecular weight markers (M; Precision Plus Protein Dual Color Standards, <t>Bio‐Rad)</t> are indicated to the right. Middle panels: Total densitometric profile of fluorescent substrate was determined for each lane. Lower panels: Coomassie blue staining (CBS) of mitochondrial proteins was used as a loading control. (D) Chemical structure of DA29 (ClpP activator) and DA28 (negative control). (E) Effect of ClpP chemoactivation by DA29 on casein hydrolysis in the PAGE‐based fluorescent assay; DA28 was used as a negative control. For description, see panel C.
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    Targeted activation of endogenous mitochondrial ClpP. (A) Western blots of mitochondrial lysates (30 μg) from wild‐type and ClpP‐KO HEK293T cells were probed with antibodies against ClpP and α‐tubulin, the latter being used as a loading control. (B) Analysis of ClpP and ClpX assembly was carried out in wild‐type HEK293T mitochondria through isokinetic sucrose gradient centrifugation. Mitochondrial lysate (0.65 mg) was centrifuged on a 10%–30% gradient using the SW 60Ti rotor at 40 000 rpm for 2 h and 15 min at 4°C. Small and large mitoribosomal subunits were used as a reference of migration and were immunodecorated by antibodies against marker proteins of the mt‐SSU (mS29; HUGO protein name: DAP3) and the mt‐LSU (uL13m; HUGO protein name: MRPL13). (C) Effect of ONC201‐mediated ClpP activation on casein hydrolysis in the PAGE‐based fluorescent assay. Mitochondrial lysates (50 μg) from wild‐type and KO cells were used as a source of ClpP activity, and incubated with ONC201 at a final concentration of 100 μM. Upper panels: Milk casein isoforms (α, β and k) are depicted to the left; molecular weight markers (M; Precision Plus Protein Dual Color Standards, <t>Bio‐Rad)</t> are indicated to the right. Middle panels: Total densitometric profile of fluorescent substrate was determined for each lane. Lower panels: Coomassie blue staining (CBS) of mitochondrial proteins was used as a loading control. (D) Chemical structure of DA29 (ClpP activator) and DA28 (negative control). (E) Effect of ClpP chemoactivation by DA29 on casein hydrolysis in the PAGE‐based fluorescent assay; DA28 was used as a negative control. For description, see panel C.
    4x Laemmli Buffer, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Targeted activation of endogenous mitochondrial ClpP. (A) Western blots of mitochondrial lysates (30 μg) from wild‐type and ClpP‐KO HEK293T cells were probed with antibodies against ClpP and α‐tubulin, the latter being used as a loading control. (B) Analysis of ClpP and ClpX assembly was carried out in wild‐type HEK293T mitochondria through isokinetic sucrose gradient centrifugation. Mitochondrial lysate (0.65 mg) was centrifuged on a 10%–30% gradient using the SW 60Ti rotor at 40 000 rpm for 2 h and 15 min at 4°C. Small and large mitoribosomal subunits were used as a reference of migration and were immunodecorated by antibodies against marker proteins of the mt‐SSU (mS29; HUGO protein name: DAP3) and the mt‐LSU (uL13m; HUGO protein name: MRPL13). (C) Effect of ONC201‐mediated ClpP activation on casein hydrolysis in the PAGE‐based fluorescent assay. Mitochondrial lysates (50 μg) from wild‐type and KO cells were used as a source of ClpP activity, and incubated with ONC201 at a final concentration of 100 μM. Upper panels: Milk casein isoforms (α, β and k) are depicted to the left; molecular weight markers (M; Precision Plus Protein Dual Color Standards, Bio‐Rad) are indicated to the right. Middle panels: Total densitometric profile of fluorescent substrate was determined for each lane. Lower panels: Coomassie blue staining (CBS) of mitochondrial proteins was used as a loading control. (D) Chemical structure of DA29 (ClpP activator) and DA28 (negative control). (E) Effect of ClpP chemoactivation by DA29 on casein hydrolysis in the PAGE‐based fluorescent assay; DA28 was used as a negative control. For description, see panel C.

    Journal: The FASEB Journal

    Article Title: Elucidating the Molecular Impact of DA29‐Induced ClpP Chemoactivation on Mitochondrial Function

    doi: 10.1096/fj.202603398RR

    Figure Lengend Snippet: Targeted activation of endogenous mitochondrial ClpP. (A) Western blots of mitochondrial lysates (30 μg) from wild‐type and ClpP‐KO HEK293T cells were probed with antibodies against ClpP and α‐tubulin, the latter being used as a loading control. (B) Analysis of ClpP and ClpX assembly was carried out in wild‐type HEK293T mitochondria through isokinetic sucrose gradient centrifugation. Mitochondrial lysate (0.65 mg) was centrifuged on a 10%–30% gradient using the SW 60Ti rotor at 40 000 rpm for 2 h and 15 min at 4°C. Small and large mitoribosomal subunits were used as a reference of migration and were immunodecorated by antibodies against marker proteins of the mt‐SSU (mS29; HUGO protein name: DAP3) and the mt‐LSU (uL13m; HUGO protein name: MRPL13). (C) Effect of ONC201‐mediated ClpP activation on casein hydrolysis in the PAGE‐based fluorescent assay. Mitochondrial lysates (50 μg) from wild‐type and KO cells were used as a source of ClpP activity, and incubated with ONC201 at a final concentration of 100 μM. Upper panels: Milk casein isoforms (α, β and k) are depicted to the left; molecular weight markers (M; Precision Plus Protein Dual Color Standards, Bio‐Rad) are indicated to the right. Middle panels: Total densitometric profile of fluorescent substrate was determined for each lane. Lower panels: Coomassie blue staining (CBS) of mitochondrial proteins was used as a loading control. (D) Chemical structure of DA29 (ClpP activator) and DA28 (negative control). (E) Effect of ClpP chemoactivation by DA29 on casein hydrolysis in the PAGE‐based fluorescent assay; DA28 was used as a negative control. For description, see panel C.

    Article Snippet: Whole‐cell proteins (20‐30 μg), mitochondrial lysates (30 μg) or sucrose gradient fractions (10 μL) were denatured in 4X Laemmli sample buffer (Bio‐Rad) complemented with 10% 2‐mercaptoethanol, separated by SDS‐PAGE and subjected to immunoblotting as described in [ ], using the following primary antibodies: ClpP (#A306224, Antibodies.com), ClpX (#ab168338, Abcam), total OXPHOS cocktail (#ab110413, Abcam), HA‐tag (#3724, Cell Signaling Technology), uL13m (#16241–1‐AP, Proteintech), VDAC1 (#ab186321, Abcam), mS29 (# MA141279 , Thermo Fisher Scientific), α‐tubulin (#T6074, Sigma‐Aldrich) and β‐actin (#A2066, Sigma‐Aldrich).

    Techniques: Activation Assay, Western Blot, Control, Analysis, Gradient Centrifugation, Migration, Marker, Fluorescence, Activity Assay, Incubation, Concentration Assay, Molecular Weight, Staining, Negative Control