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phospho-MLKL (Ser345) Recombinant Antibody

4% Paraformaldehyde-fixed L-929\uff08treated with 20 ng\/ml TNF alpha, 100 nM Smac mimetic, and 20 \u00b5M z-VAD for 8 h\uff09 (M) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (phospho-MLKL (Ser345)) monoclonal Antibody, unconjugated (bsm-54104R) 1:100, 90 min at 37\u00b0C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-40295G-FITC) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
4% Paraformaldehyde-fixed L-929\uff08treated with 20 ng\/ml TNF alpha, 100 nM Smac mimetic, and 20 \u00b5M z-VAD for 8 h\uff09 (M) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (phospho-MLKL (Ser345)) monoclonal Antibody, unconjugated (bsm-54104R) 1:100, 90 min at 37\u00b0C; followed by conjugated Goat Anti-Rabbit IgG antibody (green, bs-40295G-FITC) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.
L-929 (M) cells were treated with or without Z-VAD (20 \u03bcM) for 30 min, mTNF-\u03b1 (20 ng\/ml) for 4 h, SM-164 (100 nM) for 4 h, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with phospho-MLKL (Ser345) monoclonal antibody, unconjugated (bsm-54104R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
L-929 (M) cells were treated with or without Z-VAD (20 \u03bcM) for 30 min, mTNF-\u03b1 (20 ng\/ml) for 4 h, SM-164 (100 nM) for 4 h, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with phospho-MLKL (Ser345) monoclonal antibody, unconjugated (bsm-54104R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
Bioss unconjugated antibody vial

Applications

  • WB
  • IP
  • ICC/IF

Reactivity

  • Mouse
Overview
Catalog # bsm-54104r-100uL
Product Name phospho-MLKL (Ser345) Recombinant Antibody
Applications
WB IP ICC/IF
Reactivity Mouse
Specifications
Conjugation Unconjugated
Host Rabbit
Source Synthetic phospho-peptide corresponding to residues surrounding Ser345 of human MLKL
Modification Site Ser345
Clonality Recombinant
Isotype IgG
Concentration Lot dependent
Purification Purified by Protein A.
Storage Buffer 0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol
Storage Condition Shipped at 4°. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID 74568
Swiss Prot Q9D2Y4
Synonyms Phospho-MLKL (S345); pMLKL Ser345; Mixed lineage kinase domain-like protein, Mlkl
Background Pseudokinase that plays a key role in TNF-induced necroptosis, a programmed cell death process. Activated following phosphorylation by RIPK3, leading to homotrimerization, localization to the plasma membrane and execution of programmed necrosis characterized by calcium influx and plasma membrane damage. Does not have protein kinase activity.
Application Dilution
WB 1:300-5000
IP 1-2ug
ICC/IF 1:50-100

Frequently asked questions

Which applications is phospho-MLKL (Ser345) Recombinant Antibody (bsm-54104R) validated for, and at what dilution?

bsm-54104R is validated for WB (1:300-5000); IP (1-2ug); ICC/IF (1:50-100). These are recommended starting dilutions; optimal dilutions should be determined by the end user.

Which species does phospho-MLKL (Ser345) Recombinant Antibody (bsm-54104R) react with?

Tested reactivity: Mouse.

How should bsm-54104R be stored?

Shipped at 4°. Store at -20°C for one year. Avoid repeated freeze/thaw cycles. Storage buffer: 0.01M TBS (pH 7.4), 1% BSA, 0.02% Proclin 300, and 50% Glycerol

What immunogen was used to raise phospho-MLKL (Ser345) Recombinant Antibody (bsm-54104R)?

Synthetic phospho-peptide corresponding to residues surrounding Ser345 of human MLKL

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