DAPK1 Monoclonal antibody

DAPK1 Monoclonal Antibody for WB, IHC, IF/ICC, IF-P, FC (Intra), ELISA

Host / Isotype

Mouse / IgG1

Reactivity

human, mouse, rat

Applications

WB, IHC, IF/ICC, IF-P, FC (Intra), ELISA

Conjugate

Unconjugated

Publications

2

CloneNo.

1E2F9

Cat no : 67815-1-Ig

Synonyms

EC:2.7.11.1, Death-associated protein kinase 1, DAPK, DAP kinase 1, 1E2F9

---

Validation Data Gallery

• 

  WB analysis using 67815-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 67815-1-Ig (DAPK1 antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours. The membrane was stripped and reblotted with HRP-conjugated Lamin B1 Monoclonal antibody (HRP-66095) as loading control.

• 

  IHC staining of human breast cancer using 67815-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 67815-1-Ig (DAPK1 antibody) at dilution of 1:400 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human placenta using 67815-1-Ig

  Immunohistochemical analysis of paraffin-embedded human placenta tissue slide using 67815-1-Ig (DAPK1 antibody) at dilution of 1:2000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human stomach cancer using 67815-1-Ig

  Immunohistochemical analysis of paraffin-embedded human stomach cancer tissue slide using 67815-1-Ig (DAPK1 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse skin using 67815-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse skin tissue slide using 67815-1-Ig (DAPK1 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of mouse small intestine using 67815-1-Ig

  Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue slide using 67815-1-Ig (DAPK1 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of rat small intestine using 67815-1-Ig

  Immunohistochemical analysis of paraffin-embedded rat small intestine tissue slide using 67815-1-Ig (DAPK1 antibody) at dilution of 1:1000 (under 10x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IHC staining of human breast cancer using 67815-1-Ig

  Immunohistochemical analysis of paraffin-embedded human breast cancer tissue slide using 67815-1-Ig (DAPK1 antibody) at dilution of 1:400 (under 40x lens). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0).

• 

  IF Staining of human breast cancer using 67815-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed human breast cancer tissue using DAPK1 antibody (67815-1-Ig, Clone: 1E2F9 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

• 

  IF Staining of HT-1376 using 67815-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed HT-1376 cells using DAPK1 antibody (67815-1-Ig, Clone: 1E2F9 ) at dilution of 1:1000 and Multi-rAb CoraLite® Plus 594-Goat Anti-Mouse Recombinant Secondary Antibody (H+L) (Cat.NO. RGAM004 ).

• 

  IF Staining of HCT 116 using 67815-1-Ig

  Immunofluorescent analysis of (-20°C Ethanol) fixed HCT 116 cells using DAPK1 antibody (67815-1-Ig, Clone: 1E2F9 ) at dilution of 1:800 and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L).

• 

  FC experiment of HCT 116 using 67815-1-Ig

  1X10^6 HCT 116 cells were intracellularly stained with 0.4 ug Anti-Human DAPK1 (67815-1-Ig, Clone:1E2F9) and CoraLite®488-Conjugated Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG1 Isotype Control (MOPC-21) (65124-1-Ig, Clone: MOPC-21) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

---

Tested Applications

• Positive WB detected in: LNCaP cells, A549 cells, HeLa cells, HepG2 cells, K-562 cells, HSC-T6 cells, PC-12 cells, NIH/3T3 cells, 4T1 cells
• Positive IHC detected in: human breast cancer tissue, human placenta tissue, human stomach cancer tissue, mouse skin tissue, mouse small intestine tissue, rat small intestine tissue; Note: suggested antigen retrieval with TE buffer pH 9.0; (*) Alternatively, antigen retrieval may be performed with citrate buffer pH 6.0
• Positive IF-P detected in: human breast cancer tissue
• Positive IF/ICC detected in: HCT 116 cells, HT-1376 cells
• Positive FC (Intra) detected in: HCT 116 cells

Recommended dilution

• Western Blot (WB): WB : 1:1000-1:4000
• Immunohistochemistry (IHC): IHC : 1:200-1:800
• Immunofluorescence (IF)-P: IF-P : 1:200-1:800
• Immunofluorescence (IF)/ICC: IF/ICC : 1:400-1:1600
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 1 publications below
• IF: See 1 publications below

Product Information

67815-1-Ig targets DAPK1 in WB, IHC, IF/ICC, IF-P, FC (Intra), ELISA applications and shows reactivity with human, mouse, rat samples.

• Tested Reactivity: human, mouse, rat
• Cited Reactivity: mouse
• Host / Isotype: Mouse / IgG1
• Class: Monoclonal
• Type: Antibody
• Immunogen: DAPK1 fusion protein Ag29838
• Full Name: death-associated protein kinase 1
• Calculated Molecular Weight: 1430 aa, 160 kDa
• Observed Molecular Weight: 160 kDa
• GenBank Accession Number: BC113660
• Gene Symbol: DAPK1
• Gene ID (NCBI): 1612
• RRID: AB_2918578
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein G purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

DAPK1(Death-associated protein kinase 1) is a stress-activated tumor suppressor protein that plays a role in both proapoptotic or antiapoptotic signal transduction pathways. Loss of DAPK1 expression is associated with a selective advantage fortumor cells to resist apoptotic stimuli, allowing them to separate from the original tumor; from this point of view, DAPK1 could be considered a tumor metastases inhibitor gene(PMID:17319784).

Publications

Species	Application	Title
mouse	IF	Cell Commun Signal; P38-DAPK1 axis regulated LC3-associated phagocytosis (LAP) of microglia in an in vitro subarachnoid hemorrhage model; Authors - Xiang-Xin Chen
mouse	WB	Cell Death Differ; The PM20D1-NADA pathway protects against Parkinson's disease; Authors - Yunying Yang