Invitrogen

ZPK Polyclonal Antibody

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7 Published Figures

View all (14) ZPK antibodies

Questions & Answers

Questions & Answers

ZPK Antibody in Immunohistochemistry (Paraffin) (IHC (P))

ZPK Antibody in Immunohistochemistry (Paraffin) (IHC (P))

FIGURE: 1 / 14

ZPK Antibody (PA5-32173) in IHC (P)

Immunohistochemical analysis of paraffin-embedded human breast cancer, using MAP3K12 (Product # PA5-32173) antibody at 1:250 dilution.

ZPK Antibody in Immunohistochemistry (Paraffin) (IHC (P))

ZPK Antibody in Immunohistochemistry (Frozen) (IHC (F))

ZPK Antibody in Western Blot (WB)

ZPK Antibody in Western Blot (WB)

ZPK Antibody in Western Blot (WB)

ZPK Antibody in Western Blot (WB)

ZPK Antibody in Western Blot, Immunohistochemistry (WB, IHC)

ZPK Antibody in Western Blot, Immunoprecipitation (WB, IP)

ZPK Antibody in Western Blot (WB)

ZPK Antibody in Western Blot (WB)

ZPK Antibody in Western Blot (WB)

ZPK Antibody in Western Blot (WB)

ZPK Antibody in Western Blot (WB)

Product Image Gallery

ZPK Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemical analysis of paraffin-embedded human breast cancer, using MAP3K12 (Product # PA5-32173) antibody at 1:250 dilution.

ZPK Antibody in Immunohistochemistry (Frozen) (IHC (F)) — Immunohistochemistry (Frozen) analysis of ZPK was performed in frozen sectioned E13.5 Rat brain tissue using ZPK Polyclonal Antibody (Product # PA5-32173) at a dilution of 1:250 (Green). Red: beta Tubulin 3/ TUJ1, a mature neuron marker, stained by beta Tubulin 3/ TUJ1 antibody diluted at 1:500. Blue: Fluoroshield with DAPI.

ZPK Antibody in Western Blot (WB) — Western Blot analysis of ZPK was performed by separating 20 µg of mouse embryonic stem cell whole cell lysates by 7.5 % SDS-PAGE. Proteins were transferred to a membrane and probed with a ZPK Polyclonal Antibody (Product # PA5-32173) at a dilution of 1:1000.

ZPK Antibody in Western Blot (WB) — Western Blot using ZPK Polyclonal Antibody (Product # PA5-32173). Sample (50 µg of whole cell lysate). Lane A: mouse brain. 7.5% SDS PAGE. ZPK Polyclonal Antibody (Product # PA5-32173) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

ZPK Antibody in Western Blot (WB) — Western Blot analysis of ZPK was performed by separating 30 µg of various whole cell extracts by 10% SDS-PAGE. Proteins were transferred to a membrane and probed with a ZPK Polyclonal Antibody (Product # PA5-32173) at a dilution of 1:1000 and a HRP-conjugated anti-rabbit IgG secondary antibody.

ZPK Antibody in Western Blot (WB) — Western Blot analysis of ZPK was performed by separating 50 µg of Rat tissue extracts by 7.5% SDS-PAGE. Proteins were transferred to a membrane and probed with a ZPK Polyclonal Antibody (Product # PA5-32173) at a dilution of 1:1000.

ZPK Antibody in Western Blot, Immunohistochemistry (WB, IHC) — Figure 5 FKBP8 mediates ubiquitin-dependent DLK degradation. A , Western blot analysis for the expression of DLK, its K271R mutant, and FKBP8 in HEK293T cells. The numbers indicate the normalized relative intensity. B , statistical analysis of (A) (n = 3 for each condition; ** p < 0.01 by t test; mean +- S.E.M.). C , Western blot analysis for the immunoprecipitation assays of FKBP8 with DLK and the K271R mutant. Dual leucine zipper kinase, DLK K271R, and FLAG-epitope-tagged FKBP8 were expressed in HEK293T cells. The protein lysates were immunoprecipitated using an anti-FLAG antibody followed by SDS-PAGE analysis. D , Western blot analysis for validating DLK levels by the expression of FKBP8 with or without MG-132 treatment in primary cultured embryonic DRG neurons. Embryonic DRG neurons were incubated with or without 10 muM MG-132 treatment for 6 h and lysed for Western blot analysis. E , primary cultured embryonic DRG neurons were infected by lentivirus of FLAG-FKBP8 with or without Atg5 -shRNA at DIV2 and lysed for Western blot analysis. F , relative expression of Atg5 mRNA analyzed via RT-qPCR (FC, fold change; n = 3; mean +- SEM; ** p < 0.01 by t test). G , sciatic nerves from AAV-control- or AAV-FKBP8-injected mice were ligated and dissected at 24 h after injury. Longitudinal sections were immunostained with the anti-DLK antibody and TUJI antibody. Yellow dotted arrows indicate the injury site. The scale bar represents 200 mum. H , relative normalized intensity of DLK wa

ZPK Antibody in Western Blot, Immunoprecipitation (WB, IP) — Figure 3 FKBPL and FKBP8 induced lysosome-dependent DLK degradation. A , comparative analysis of relative expression levels in mouse L4,5 dorsal root ganglion (DRG) tissues, sciatic nerve tissues, and cultured embryonic DRG neurons (, , , ). Red and blue circles indicate Illumina short-read sequencing and Oxford Nanopore direct RNA long-read sequencing, respectively. Circle sizes indicate relative levels of microarray data from cultured embryonic DRG neurons. B , Western blot analysis for the expression of DLK with FKBPs (N; null vector, L; FKBPL, 3; FKBP3, 4; FKBP4, 8; FKBP8, 12; FKBP12, 14; FKBP14, 15; FKBP15). The number indicates normalized relative intensity. Dual leucine zipper kinase and FLAG-epitope-tagged FKBP protein family members were expressed in HEK293T cells and subjected to SDS-PAGE. C , Western blot analysis for the immunoprecipitation of DLK with mouse (m) and human (h) FKBP4/8 that was overexpressed in HEK293T cells. Empty arrowhead , non-specific band; blue arrowhead , FKBP4; red arrowhead , FKBP8. D , Western blot analysis for the expression of DLK and FKBPL/4/8 expressed in HEK293T cells with or without bafilomycin A1 treatment. The numbers indicate the normalized relative intensity. E , Western blot analysis of DLK protein levels under Fkbp8 knockdown (sh Fkbp8 ) by lentiviral delivery in primary cultured embryonic DRG neurons. The numbers indicate the normalized relative intensity. F , statistical analysis of ( E ) (FC, fold change; n = 3 for each cond

ZPK Antibody in Western Blot (WB) — Fig. 4 GSK2334470, a chemical inhibitor of PDK1, promotes axon regeneration in vitro. a In vitro axon regeneration assays. Mouse embryonic DRG neurons were re-plated in GSK2334470 (1 muM or 5 muM as indicated) or a vehicle (control) containing culture medium, fixed, and immunostained with TUJ1 antibody. Scale bar, 100 mum. b Average length of the regenerating axons from ( a ) (Mean +- S.E.M. ; n = 151, 156, and 161 cells for each condition; ** p < 0.01 from a t -test). c Western blot analysis of protein lysates from control (0, vehicle only) or GSK2334470-treated embryonic DRG neuron cultures. Numbers indicate the relative intensities

ZPK Antibody in Western Blot (WB) — Fig. 6 PDK1 knockdown leads to downregulation of the DLK/c-jun pathway. a Western blot analysis of protein lysates from control or PDK1-knockdown embryonic DRG neurons. b Relative intensity of ( a ) (Mean +- S.E.M. ; n = 4 for each condition; *** p < 0.001, ** p < 0.01 from a t -test). c Relative mRNA levels measured using quantitative RT-PCR from cultured embryonic DRG neurons (Mean +- S.D. ; n = 6; *** p < 0.001 from a t -test)

ZPK Antibody in Western Blot (WB) — Figure 4 DLK is constitutively active in all the brain regions of adult mouse brain but has brain region specific scaffolding proteins. ( a ) Representative immunoblots of DLK from the homogenates obtained from nTg and Tg forebrain of 2, 4, 6, and 8 months old female mice. ( b ) Quantification of the immunoblots of DLK/GAPDH from the nTg and Tg forebrain homogenates from 2, 4, 6, and 8 months old female mice, plotted as mean fold change relative to nTg group of respective age cohort. Unpaired t -test. + p < 0.05, ++ p < 0.01, n = 6. ( c ) Immunoblots showing differential expression of DLK signaling components: p-MKK4, MKK4, JNK1, JNK2, JNK3, p-JNK, JNK, p-c-Jun, and c-Jun from cerebellum (Cbm) and forebrain (Fore) from 3-4 months old male WT mice. ( d ) Representative immunoblots of p-MKK4 and MKK4 in homogenates from nTg cerebellum (nTg Cbm), nTg forebrain (nTg Fore), and Tg forebrain (Tg Fore) of 7-8 months old female mice treated with 50 mg/kg of DLKi or vehicle QD for 7 days. ( e ) Quantification of the immunoblots of p-MKK4/GAPDH and MKK4/GAPDH from homogenates obtained from nTg cerebellum, nTg forebrain, and Tg forebrain of mice treated with vehicle or DLKi, plotted as mean fold change relative to vehicle group in the respective brain region. Unpaired t -test. ** p < 0.01, n = 6. ( f ) Immunoblots showing differential expression of DLK scaffolding proteins: JIP1, JIP2, JIP3, and POSH from cerebellum (Cbm) and forebrain (Fore) from 3-4 months old male WT mice.

ZPK Antibody in Western Blot (WB) — Figure 9 Acute pharmacological inhibition of DLK is sufficient to upregulate IGFBP5 in uninjured mouse cerebellum. ( a ) Schematic representation of study timeline; 3-4 months old male WT mice were treated with a single oral dose of 50 mg/kg of DLKi or vehicle QD. The study was terminated approximately 2, 4, 8, 12, or 24 h after single dose. ( b ) Representative immunoblots of p-c-Jun, c-Jun, IGFBP5, p-IGF1R, and IGF1R from cerebellar homogenates of WT mice treated with a single dose of vehicle or DLKi for either 2, 4, 8, 12, or 24 h. ( c ) Quantification of the immunoblots of p-c-Jun/GAPDH, c-Jun/GAPDH, IGFBP5/GAPDH, and p-IGF1R/IGF1R from cerebellar homogenates of WT mice treated with single dose of vehicle or DLKi for either 2, 4, 8, 12, or 24 h, plotted as mean fold change relative to vehicle group at respective time-point. Unpaired t -test. * p < 0.05, ** p < 0.01, n = 5.

ZPK Antibody in Western Blot (WB) — Published figure using ZPK polyclonal antibody (Product # PA5-32173) in Western Blot

ZPK Antibody — Figure 4 Activation of the DLK/JIP3-JNK pathway is required for Phase I gene expression during HSV reactivation. ( A. ) ICP27 mRNA levels at different time points following reactivation stimulated by LY294002 as determined by RT-qPCR. The relative copy number of ICP27 mRNA was normalized to the relative GAPDH copy number in the same sample. The two previously characterized phases of HSV reactivation are highlighted (Means +/- SEM, n=3). Reactivation was carried out in the presence of WAY-150138. ( B. ) ICP27 mRNA levels at 18h post-reactivation in the presence and absence of SP600125. ( C-E. ) ICP27 mRNA levels at 18h post-reactivation following lentivirus shRNA-mediated depletion of DLK or JIP3, or infection with the control lentivirus (pLKO.1). Western blots of DLK and JIP3 protein levels with the relative levels of DLK and JIP3 normalized to beta-actin (below). B-E represent the mean +/- SEM n>=5. See also figure S3 . {KD}

Product Details

PA5-32173

Applications

Tested Dilution

Publications

Western Blot (WB)

1:500-1:3,000

View 6 publications 6 publications

Immunohistochemistry (IHC)

-

View 1 publication 1 publication

Immunohistochemistry (Paraffin) (IHC (P))

1:100-1:1,000

-

Immunohistochemistry (Frozen) (IHC (F))

1:100-1:1,000

-

Immunoprecipitation (IP)

-

View 1 publication 1 publication

Product Specifications

Species Reactivity

Human, Mouse, Rat

Published species

Mouse

Host/Isotype

Rabbit / IgG

Class

Polyclonal

Type

Antibody

Immunogen

Recombinant protein encompassing a sequence within the center region of mouse DLK/MAP3K12. The exact sequence is proprietary.

Conjugate

Unconjugated Unconjugated Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Antigen affinity chromatography

Storage buffer

PBS, pH 7, with 1% BSA, 20% glycerol

Contains

0.01% thimerosal

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Shipping conditions

Ambient (domestic); Wet ice (international)

AB_2549646

Product Specific Information

Recommended positive controls: Neuro 2A, C8D30, NIH-3T3, Raw264.7, C2C12, Mouse brain, mouse embryonic stem cell, rat brain.

Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Target Information

ZPK is a member of serine/threonine protein kinase family. This kinase contains a leucine-zipper domain, and is predominately expressed in neuronal cells. The phosphorylation state of this kinase in synaptic terminals was shown to be regulated by membrane depolarization via calcineurin. This kinase forms heterodimers with leucine zipper containing transcription factors, such as cAMP responsive element binding protein (CREB) and MYC, and thus may play a regulatory role in PKA or retinoic acid induced neuronal differentiation.

⚠WARNING: This product can expose you to chemicals including mercury, which is known to the State of California to cause birth defects or other reproductive harm. For more information go to www.P65Warnings.ca.gov.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

FK506-binding protein-like and FK506-binding protein 8 regulate dual leucine zipper kinase degradation and neuronal responses to axon injury.

The Journal of biological chemistry

Lee B,Oh Y,Cho E,DiAntonio A,Cavalli V,Shin JE,Choi HW,Cho Y

PA5-32173 was used in Immunoprecipitation, Immunohistochemistry to investigate the molecular mechanism of DLK function in response to neuronal injury.

Tue Mar 01 00:00:00 UTC 2022

FK506-binding protein-like and FK506-binding protein 8 regulate dual leucine zipper kinase degradation and neuronal responses to axon injury.

The Journal of biological chemistry

Lee B,Oh Y,Cho E,DiAntonio A,Cavalli V,Shin JE,Choi HW,Cho Y

PA5-32173 was used in Immunoprecipitation, Immunohistochemistry to investigate the molecular mechanism of DLK function in response to neuronal injury.

Tue Mar 01 00:00:00 UTC 2022

FK506-binding protein-like and FK506-binding protein 8 regulate dual leucine zipper kinase degradation and neuronal responses to axon injury.

The Journal of biological chemistry

Lee B,Oh Y,Cho E,DiAntonio A,Cavalli V,Shin JE,Choi HW,Cho Y

PA5-32173 was used in Immunoprecipitation, Immunohistochemistry to investigate the molecular mechanism of DLK function in response to neuronal injury.

Tue Mar 01 00:00:00 UTC 2022

FK506-binding protein-like and FK506-binding protein 8 regulate dual leucine zipper kinase degradation and neuronal responses to axon injury.

The Journal of biological chemistry

Lee B,Oh Y,Cho E,DiAntonio A,Cavalli V,Shin JE,Choi HW,Cho Y

PA5-32173 was used in Immunoprecipitation, Immunohistochemistry to investigate the molecular mechanism of DLK function in response to neuronal injury.

Tue Mar 01 00:00:00 UTC 2022

FK506-binding protein-like and FK506-binding protein 8 regulate dual leucine zipper kinase degradation and neuronal responses to axon injury.

The Journal of biological chemistry

Lee B,Oh Y,Cho E,DiAntonio A,Cavalli V,Shin JE,Choi HW,Cho Y

PA5-32173 was used in Immunoprecipitation, Immunohistochemistry to investigate the molecular mechanism of DLK function in response to neuronal injury.

Tue Mar 01 00:00:00 UTC 2022

PDK1 is a negative regulator of axon regeneration.

Molecular brain

Kim H,Lee J,Cho Y

PA5-32173 was used in Western Blot to present PDK1 as a new player to negatively regulate axon regeneration and as a potential target in the development of therapeutic applications.

Fri Feb 12 00:00:00 UTC 2021

PDK1 is a negative regulator of axon regeneration.

Molecular brain

Kim H,Lee J,Cho Y

PA5-32173 was used in Western Blot to present PDK1 as a new player to negatively regulate axon regeneration and as a potential target in the development of therapeutic applications.

Fri Feb 12 00:00:00 UTC 2021

Dual Leucine Zipper Kinase Is Constitutively Active in the Adult Mouse Brain and Has Both Stress-Induced and Homeostatic Functions.

International journal of molecular sciences

Goodwani S,Fernandez C,Acton PJ,Buggia-Prevot V,McReynolds ML,Ma J,Hu CH,Hamby ME,Jiang Y,Le K,Soth MJ,Jones P,Ray WJ

Published figure using ZPK polyclonal antibody (Product # PA5-32173) in Western Blot

Thu Jul 09 00:00:00 UTC 2020

Neuronal hyperexcitability is a DLK-dependent trigger of herpes simplex virus reactivation that can be induced by IL-1.

eLife

Cuddy SR,Schinlever AR,Dochnal S,Seegren PV,Suzich J,Kundu P,Downs TK,Farah M,Desai BN,Boutell C,Cliffe AR

PA5-32173 was used in Western Blotting to investigate the stimuli that triggers HSV-1 reactivation in a mouse model.

Tue Dec 22 00:00:00 UTC 2020

Coupled Control of Distal Axon Integrity and Somal Responses to Axonal Damage by the Palmitoyl Acyltransferase ZDHHC17.

Cell reports

Niu J,Sanders SS,Jeong HK,Holland SM,Sun Y,Collura KM,Hernandez LM,Huang H,Hayden MR,Smith GM,Hu Y,Jin Y,Thomas GM

PA5-32173 was used in Western Blot to demonstrate that two palmitoylation-dependent pathways mediate the control of somal and distal axon integrity.

Tue Nov 17 00:00:00 UTC 2020

Neuronal Stress Pathway Mediating a Histone Methyl/Phospho Switch Is Required for Herpes Simplex Virus Reactivation.

Cell host & microbe

Cliffe AR,Arbuckle JH,Vogel JL,Geden MJ,Rothbart SB,Cusack CL,Strahl BD,Kristie TM,Deshmukh M

Published figure using ZPK polyclonal antibody (Product # PA5-32173) in Western Blot

Wed Dec 09 00:00:00 UTC 2015

Bioinformatics

Protein Aliases: DLK; Dual leucine zipper bearing kinase; dual leucine zipper kinase; dual leucine zipper kinase DLK; leucine zipper protein kinase; Leucine-zipper protein kinase; M3KC; MAPK-upstream kinase; Mitogen activated protein kinase 12 (Zipper (leucine) protein kinase); mitogen activated protein kinase kinase kinase 12; Mitogen-activated protein kinase kinase kinase 12; Mixed lineage kinase; MUK; protein kinase MUK; Zipper (leucine) protein kinase; ZPK

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Gene Aliases: DLK; MAP3K12; MEKK12; MUK; PK; ZPK; ZPKP1

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UniProt ID: (Human) Q12852, (Rat) Q63796, (Mouse) Q60700

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Entrez Gene ID: (Human) 7786, (Rat) 25579, (Mouse) 26404

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protein kinase activity protein serine/threonine kinase activity MAP kinase kinase kinase activity protein binding ATP binding protein kinase binding protein homodimerization activity nucleotide binding kinase activity transferase activity transferase activity, transferring phosphorus-containing groups

activation of MAPKK activity protein phosphorylation JNK cascade histone phosphorylation peptidyl-serine phosphorylation peptidyl-threonine phosphorylation protein autophosphorylation phosphorylation mitophagy in response to mitochondrial depolarization intracellular signal transduction negative regulation of motor neuron apoptotic process

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