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phospho-MLKL (Ser345)

L-929 (M) cells were treated with or without mTNF alpha (20 ng\/ml) Smac mimetic (100 nM) and z-VAD (20 \u00b5M) for 8 h, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with phospho-MLKL (Ser345) polyclonal antibody, unconjugated (bs-25447R) at 1:500 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 mTNF alpha (20 ng\/ml) Smac mimetic (100 nM) and z-VAD (20 \u00b5M) for 8 h, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with phospho-MLKL (Ser345) polyclonal antibody, unconjugated (bs-25447R) at 1:500 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.
4% Paraformaldehyde-fixed L-929 (treated with 20 ng\/ml TNF alpha, 100 nM Smac mimetic, and 20 \u00b5M z-VAD for 8 h) (M) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (phospho-MLKL (Ser345)) polyclonal Antibody, unconjugated (bs-25447R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) 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 (treated with 20 ng\/ml TNF alpha, 100 nM Smac mimetic, and 20 \u00b5M z-VAD for 8 h) (M) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (phospho-MLKL (Ser345)) polyclonal Antibody, unconjugated (bs-25447R) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Rabbit IgG antibody (green, bs-60295G-BF488) 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.
Bioss unconjugated antibody vial

Applications

  • WB

Reactivity

  • Mouse

Predicted Reactivity

  • Mouse
Overview
Catalog # bs-25447r
Product Name phospho-MLKL (Ser345)
Applications
WB
Reactivity Mouse
Predicted Reactivity Mouse
Specifications
Conjugation Unconjugated
Host Rabbit
Source KLH conjugated Synthesised phosphopeptide derived from mouse MLKL around the phosphorylation site of Ser345
Clonality Polyclonal
Isotype IgG
Concentration 1mg/ml
Purification Purified by Protein A.
Storage Buffer 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles.
Target
Gene ID 197259
Swiss Prot Q9D2Y4
Subcellular location Cytoplasm, Cell membrane
Synonyms phospho-MLKL(S345); p-MLKL(S345); MLKL(p-S345); hMLKL; Mixed lineage kinase domain like; Mixed lineage kinase domain like protein; Mixed lineage kinase domain-like protein; mixed lineage kinase domain like pseudokinase; MLKL_HUMAN.
Application Dilution
WB 1:200-1000

Frequently asked questions

Which applications is phospho-MLKL (Ser345) (bs-25447R) validated for, and at what dilution?

bs-25447R is validated for WB (1:200-1000). These are recommended starting dilutions; optimal dilutions should be determined by the end user.

Which species does phospho-MLKL (Ser345) (bs-25447R) react with?

Tested reactivity: Mouse. Predicted reactivity (expected from sequence homology but not yet tested): Mouse.

How should bs-25447R be stored?

Shipped at 4C. Store at -20C for one year. Avoid repeated freeze/thaw cycles. Storage buffer: 0.01M TBS (pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

What immunogen was used to raise phospho-MLKL (Ser345) (bs-25447R)?

KLH conjugated Synthesised phosphopeptide derived from mouse MLKL around the phosphorylation site of Ser345

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