MLKL Polyclonal Antibody

$Paraformaldehyde-fixed, paraffin embedded Human breast cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with MLKL Polyclonal Antibody, Unconjugated (bs-55131R) at 1:100 overnight at 4\u00b0C, DAB staining.$

Paraformaldehyde-fixed, paraffin embedded Human breast cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with MLKL Polyclonal Antibody, Unconjugated (bs-55131R) at 1:100 overnight at 4\u00b0C, DAB staining.

$Lane 1: Jurkat cell lysates; Lane 2: NCI-H460 cell lysates probed with MLKL Polyclonal Antibody, Unconjugated (bs-55131R) at 1:1000 dilution and 4\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.$

Lane 1: Jurkat cell lysates; Lane 2: NCI-H460 cell lysates probed with MLKL Polyclonal Antibody, Unconjugated (bs-55131R) at 1:1000 dilution and 4\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.

$Lane 1: Mouse Liver lysates probed with MLKL Polyclonal Antibody, Unconjugated (bs-55131R) at 1:1000 dilution and 4\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.$

Lane 1: Mouse Liver lysates probed with MLKL Polyclonal Antibody, Unconjugated (bs-55131R) at 1:1000 dilution and 4\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.

$Lane 1: Hela cell lysates; Lane 2: MLKL knockout (KO) HeLa cell lysates probed with MLKL Polyclonal Antibody, Unconjugated (bs-55131R) at 1:1000 dilution and 4\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.$

Lane 1: Hela cell lysates; Lane 2: MLKL knockout (KO) HeLa cell lysates probed with MLKL Polyclonal Antibody, Unconjugated (bs-55131R) at 1:1000 dilution and 4\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.

$Lane 1: Rat Testis lysates probed with MLKL Polyclonal Antibody, Unconjugated (bs-55131R) at 1:1000 dilution and 4\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.$

Lane 1: Rat Testis lysates probed with MLKL Polyclonal Antibody, Unconjugated (bs-55131R) at 1:1000 dilution and 4\u02daC overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.

$Lane 1: Human HeLa cell lysates; Lane 2: Human MCF-7 cell lysates probed with MLKL Polyclonal Antibody, Unconjugated (bs-55131R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.$

Lane 1: Human HeLa cell lysates; Lane 2: Human MCF-7 cell lysates probed with MLKL Polyclonal Antibody, Unconjugated (bs-55131R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37\u02daC.

$25 ug total protein per lane of various lysates (see on figure) probed with MLKL polyclonal antibody, unconjugated (bs-55131R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.$

25 ug total protein per lane of various lysates (see on figure) probed with MLKL polyclonal antibody, unconjugated (bs-55131R) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

Applications

WB
IHC-P
IHC-F
IF

Reactivity

Human

Overview
Catalog #	bs-55131R
Product Name	MLKL Polyclonal Antibody
Applications	WB IHC-P IHC-F IF
Reactivity	Human
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Recombinant protein of mouse MLKL
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	74568
Swiss Prot	Q9D2Y4
Subcellular location	Cytoplasm, Cell membrane
Synonyms	hMLKL; 9130019I15Rik; MLKL_HUMAN; MLKL; MLKL_MOUSE; mixed lineage kinase domain like pseudokinase; mixed lineage kinase domain-like
Background	This gene belongs to the protein kinase superfamily. The encoded protein contains a protein kinase-like domain; however, is thought to be inactive because it lacks several residues required for activity. This protein plays a critical role in tumor necrosis factor (TNF)-induced necroptosis, a programmed cell death process, via interaction with receptor-interacting protein 3 (RIP3), which is a key signaling molecule in necroptosis pathway. Inhibitor studies and knockdown of this gene inhibited TNF-induced necrosis. High levels of this protein and RIP3 are associated with inflammatory bowel disease in children. Alternatively spliced transcript variants have been described for this gene.
Application Dilution
WB	1:300-5000
IHC-P	1:200-400
IHC-F	IHC
IF