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Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Datasheet

Tested applications:WBIHC-PIF/ICCIPChIPChIP-seqRIPFCFC(Intra)ELISAMeDIPNucleotide ArrayDBFACSCoIPCUT&TagmeRIPInhibitionReactivity:Human, Mouse, Rat

ABclonal:Western blot - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Western blot analysis of lysates from C6 cells using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at 1:2600 dilution incubated overnight at 4℃. C6 cells were treated by UV (90mJ/cm2) at room temperature and recovered for 2 hours.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 30 μg per lane.
Blocking buffer: 3 % nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 60s.

ABclonal:Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Immunohistochemistry analysis of paraffin-embedded Human colon tissue, Untreated (left) and lambda phosphatase-treated (right), using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

ABclonal:Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Immunohistochemistry analysis of paraffin-embedded Mouse spleen tissue, Untreated (left) and lambda phosphatase-treated (right), using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

ABclonal:Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Immunohistochemistry analysis of paraffin-embedded Rat spleen tissue, Untreated (left) and lambda phosphatase-treated (right), using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

ABclonal:Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Immunohistochemistry analysis of paraffin-embedded Human breast cancer tissue using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

ABclonal:Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Immunohistochemistry analysis of paraffin-embedded Rat kidney tissue using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

ABclonal:Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Immunohistochemistry analysis of paraffin-embedded Mouse brain tissue using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

ABclonal:Immunofluorescence - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Confocal imaging of HeLa cells (treated with UV) and HeLa cells (untreated) using Phospho-MAP2K4-S257 Rabbit mAb (AP1491, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.

ABclonal:Immunoprecipitation - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Immunoprecipitation of Phospho-MAP2K4-S257 from 200 µg extracts of HeLa cells treated by UV (100 mJ/cm2, 4 hours) was performed using 0.5 µg of Phospho-MAP2K4-S257 Rabbit mAb (AP1491). Rabbit IgG isotype control (AC042) was used to precipitate the Control IgG sample. IP samples were eluted with 1X Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at a dilution of 1:1000.

Overview

Product namePhospho-MAP2K4-S257 Rabbit mAb
Catalog No.AP1491
Host speciesRabbit
Purification methodAffinity purification
IsotypeIgG
CloneNo.ARC66577
This gene encodes a member of the mitogen-activated protein kinase (MAPK) family. Members of this family act as an integration point for multiple biochemical signals and are involved in a wide variety of cellular processes such as proliferation, differentiation, transcription regulation, and development. They form a three-tiered signaling module composed of MAPKKKs, MAPKKs, and MAPKs. This protein is phosphorylated at serine and threonine residues by MAPKKKs and subsequently phosphorylates downstream MAPK targets at threonine and tyrosine residues. A similar protein in mouse has been reported to play a role in liver organogenesis. A pseudogene of this gene is located on the long arm of chromosome X. Alternative splicing results in multiple transcript variants.
ImmunogenA synthetic phosphorylated peptide around S257 of human MAP2K4 (NP_003001.1).
SequenceVDS(P)IA
Gene ID
Swiss Prot
SynonymsJNKK; MEK4; MKK4; SEK1; SKK1; JNKK1; SERK1; MAPKK4; PRKMK4; SAPKK1; SAPKK-1
Calculated MW44kDa
Observed MW44kDa
ReactivityHuman, Mouse, Rat
Tested applicationsWBIHC-PIF/ICCIPChIPChIP-seqRIPFCFC(Intra)ELISAMeDIPNucleotide ArrayDBFACSCoIPCUT&TagmeRIPInhibition
Recommended dilution
  • WB 1:1000 - 1:5000
  • IHC-P 1:50 - 1:200
  • IF/ICC 1:50 - 1:200
  • IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells
Storage bufferStore at -20℃. Avoid freeze / thaw cycles.
Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Key applicationWestern blotting    Immunohistochemistry    Immunofluorescence    Immunoprecipitation    
Positive samplesHeLa+UV, C6+UV
Cellular locationCytoplasm, Nucleus

Documents

Certificate of Compliance

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ABclonal:Western blot - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)}

Western blot - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Western blot analysis of lysates from C6 cells using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at 1:2600 dilution incubated overnight at 4℃. C6 cells were treated by UV (90mJ/cm2) at room temperature and recovered for 2 hours.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 30 μg per lane.
Blocking buffer: 3 % nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 60s.
ABclonal:Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)}

Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Immunohistochemistry analysis of paraffin-embedded Human colon tissue, Untreated (left) and lambda phosphatase-treated (right), using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.
ABclonal:Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)}

Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Immunohistochemistry analysis of paraffin-embedded Mouse spleen tissue, Untreated (left) and lambda phosphatase-treated (right), using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.
ABclonal:Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)}

Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Immunohistochemistry analysis of paraffin-embedded Rat spleen tissue, Untreated (left) and lambda phosphatase-treated (right), using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.
ABclonal:Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)}

Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Immunohistochemistry analysis of paraffin-embedded Human breast cancer tissue using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.
ABclonal:Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)}

Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Immunohistochemistry analysis of paraffin-embedded Rat kidney tissue using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.
ABclonal:Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)}

Immunohistochemistry - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Immunohistochemistry analysis of paraffin-embedded Mouse brain tissue using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at a dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.
ABclonal:Immunofluorescence - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)}

Immunofluorescence - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Confocal imaging of HeLa cells (treated with UV) and HeLa cells (untreated) using Phospho-MAP2K4-S257 Rabbit mAb (AP1491, dilution 1:200) followed by a further incubation with Cy3 Goat Anti-Rabbit IgG (H+L) (AS007, dilution 1:500) (Red). DAPI was used for nuclear staining (Blue). Objective: 100x.
ABclonal:Immunoprecipitation - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)}

Immunoprecipitation - Phospho-MAP2K4-S257 Rabbit mAb (AP1491)

Immunoprecipitation of Phospho-MAP2K4-S257 from 200 µg extracts of HeLa cells treated by UV (100 mJ/cm2, 4 hours) was performed using 0.5 µg of Phospho-MAP2K4-S257 Rabbit mAb (AP1491). Rabbit IgG isotype control (AC042) was used to precipitate the Control IgG sample. IP samples were eluted with 1X Laemmli Buffer. The Input lane represents 10% of the total input. Western blot analysis of immunoprecipitates was conducted using Phospho-MAP2K4-S257 Rabbit mAb (AP1491) at a dilution of 1:1000.

* For research use only. Not for therapeutic or diagnostic purposes.

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