- PA5-79716 - Invitrogen Antibodies MYLK antibody

Submitted references The Host CYP1A1-Microbiota Metabolic Axis Promotes Gut Barrier Disruption in Methicillin-Resistant Staphylococcus aureus-Induced Abdominal Sepsis.

Ma X, Jin H, Chu X, Dai W, Tang W, Zhu J, Wang F, Yang X, Li W, Liu G, Yang X, Liang H
Frontiers in microbiology 2022;13:802409

Transcriptional profiling of intervertebral disc in a post-traumatic early degeneration organ culture model.

Cui S, Zhou Z, Chen X, Wei F, Richards RG, Alini M, Grad S, Li Z
JOR spine 2021 Sep;4(3):e1146

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of MYLK in Lane 1: human SGC-7901 whole cell lysate, Lane 2: rat spleen tissue lysate, Lane 3: mouse spleen tissue lysate using 50 µg (reducing conditions) per well. Electrophoresis was performed on 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours and protein was transferred to a nitrocellulose membrane at 150mA for 50-90 minutes. Sample was blocked with 5% Non-fat Milk/TBS for 1.5 hours at room temperature, incubated with MYLK polyclonal antibody (Product # PA5-79716) at a dilution of 0.5 µg/mL (overnight at 4°C), followed by goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:10,000. Signal development was performed using a chemiluminescence (ECL) kit.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of MYLK was performed using 70% confluent log phase Caco-2 cells and Caco-2 cells treated with 10 ng/mL of IFN gamma for 18 hr followed by 2.5 ng/mL of TNF alpha for 4 hr. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with MYLK Rabbit Polyclonal Antibody (Product # PA5-79716) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green).Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image of treated cells showing cytoplasm & cytoskeletal localization. Panel e represents untreated cells with no signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of MYLK was performed using 70% confluent log phase Caco-2 cells and Caco-2 cells treated with 10 ng/mL of IFN gamma for 18 hr followed by 2.5 ng/mL of TNF alpha for 4 hr. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with MYLK Rabbit Polyclonal Antibody (Product # PA5-79716) at 1:100 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green).Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image of treated cells showing cytoplasm & cytoskeletal localization. Panel e represents untreated cells with no signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of MYLK on paraffin-embedded human rectal cancer tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with MYLK polyclonal antibody (Product# PA5-79716) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of MYLK on paraffin-embedded human placenta tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with MYLK polyclonal antibody (Product# PA5-79716) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of MYLK on paraffin-embedded rat small intestine tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with MYLK polyclonal antibody (Product# PA5-79716) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of MYLK on paraffin-embedded mouse small intestine tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with MYLK polyclonal antibody (Product# PA5-79716) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of MYLK on paraffin-embedded mouse lung tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with MYLK polyclonal antibody (Product# PA5-79716) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of MYLK on paraffin-embedded rat lung tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with MYLK polyclonal antibody (Product# PA5-79716) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of MYLK on paraffin-embedded human placenta tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with MYLK polyclonal antibody (Product# PA5-79716) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry analysis of MYLK on paraffin-embedded rat small intestine tissue. Antigen retrieval was performed using citrate buffer (pH6, epitope retrieval solution) for 20 mins. Sample was blocked using 10% goat serum, incubated with MYLK polyclonal antibody (Product# PA5-79716) with a dilution of 1 µg/mL (overnight at 4°C), and followed by biotinylated goat anti-rabbit IgG (30 minutes at 37°C). Development was performed using Streptavidin-Biotin-Complex (SABC) with DAB chromogen method.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: FIGURE 6 Microbiota-derived CAD induces gut barrier disruption through HRH4/NF-kappaB/MLCK activation. (A) Relative mRNA expression of HRH1, HRH2, HRH3, and HRH4 in the ileal epithelium from each group ( n = 3-4). (B) Immunoblot and densitometry plots of the MRSA-induced HRH4 levels in the ileal epithelium from each group ( n = 5-6). (C,D) Immunoblot and densitometry plots of MLCK (C) and P-MLC2 (D) in whole-cell lysates of IECs from each group ( n = 5-6). (E) Immunoblot and densitometry plots of tight junction proteins in the ileal epithelium from each group ( n = 6-7). (F,G) Immunoblot and densitometry plots of the p65 levels in the nuclear extracts (F) and those of P-IkappaBalpha levels in the cytosolic extracts (G) of IECs from two mice before or at 12 h p-Mi ( Cyp1a1 +/+ mice, n = 6-7; Cyp1a1 -/- mice, n = 6-7). PCNA is a nuclear loading control. (H-J) Immunoblot and densitometry plots of the p65 levels in the nuclear extracts (H) , P-IkappaBalpha levels in the cytosolic extracts (I) , and tight junction protein levels in whole-cell lysates (J) of IECs from each group ( n = 6-7). Error bars, +- SEM. Differences were analyzed using one-way ANOVA. * P < 0.05, ** P < 0.01, *** P < 0.001, n.s. not significant. # Comparison between Cyp1a1 +/+ _MRSA and Cyp1a1 -/- _MRSA groups or between JNJ7777120 - MRSA + and JNJ7777120 + MRSA + groups. # P < 0.05.

PA5-79716

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