Recombinant Anti-PYK2 antibody [YE353] (ab32571)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [YE353] to PYK2
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-PYK2 antibody [YE353]
See all PYK2 primary antibodies -
Description
Rabbit monoclonal [YE353] to PYK2 -
Host species
Rabbit -
Specificity
This antibody recognizes PYK2. It does not cross react with other FAK family members. -
Tested applications
Suitable for: WB, IHC-P, ICC/IFmore details
Unsuitable for: IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human PYK2 aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: Q14289 -
Positive control
- WB: Ramos, Jurkat and RAW264.7 cell lysates and mouse and rat brain tissue lysates. IHC-P: Human, mouse and rat cerebral cortex tissues. ICC/IF: HeLa and PC12 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
YE353 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32571 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
1/2000. Detects a band of approximately 116 kDa (predicted molecular weight: 116 kDa).
For unpurified use at 1/1000 - 1/5000. |
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IHC-P |
1/300. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/250 - 1/500. |
|
ICC/IF |
1/60.
For unpurified use at 1/100. |
Notes |
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WB
1/2000. Detects a band of approximately 116 kDa (predicted molecular weight: 116 kDa). For unpurified use at 1/1000 - 1/5000. |
IHC-P
1/300. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/250 - 1/500. |
ICC/IF
1/60. For unpurified use at 1/100. |
Target
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Function
Involved in calcium induced regulation of ion channel and activation of the map kinase signaling pathway. May represent an important signaling intermediate between neuropeptide activated receptors or neurotransmitters that increase calcium flux and the downstream signals that regulate neuronal activity. Interacts with the SH2 domain of Grb2. May phosphorylate the voltage-gated potassium channel protein Kv1.2. Its activation is highly correlated with the stimulation of c-Jun N-terminal kinase activity. Involved in osmotic stress-dependent SNCA 'Tyr-125' phosphorylation. In concert with SRC, plays an important role in osteoclastic bone resorption. Both the formation of a SRC-PTK2B complex, and SRC kinase activity are necessary for this function. The Tyr-402 phosphorylated form serves as a docking site for SRC and is important for the organization of the osteoclast actin cytoskeleton and attachment sites and for bone resorption. -
Tissue specificity
Most abundant in the brain, with highest levels in amygdala and hippocampus. Low levels in kidney. Also expressed in spleen and lymphocytes. -
Sequence similarities
Belongs to the protein kinase superfamily. Tyr protein kinase family. FAK subfamily.
Contains 1 FERM domain.
Contains 1 protein kinase domain. -
Post-translational
modificationsPhosphorylated on tyrosine residues in response to various stimuli that elevate the intracellular calcium concentration, as well as by PKC activation. Recruitment by nephrocystin to cell matrix adhesions initiates Tyr-402 phosphorylation. In monocytes, adherence to substrata is required for tyrosine phosphorylation and kinase activation. Angiotensin II, thapsigargin and L-alpha-lysophosphatidic acid (LPA) also induce autophosphorylation and increase kinase activity. -
Cellular localization
Cytoplasm. Cell membrane. Interaction with nephrocystin induces the membrane-association of the kinase. - Information by UniProt
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Database links
- Entrez Gene: 2185 Human
- Entrez Gene: 19229 Mouse
- Entrez Gene: 50646 Rat
- Omim: 601212 Human
- SwissProt: Q14289 Human
- SwissProt: Q9QVP9 Mouse
- SwissProt: P70600 Rat
- Unigene: 491322 Human
see all -
Alternative names
- CADTK antibody
- CAK-beta antibody
- CAKB antibody
see all
Images
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Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: PYK2 knockout HAP1 whole cell lysate (20 µg)
Lane 3: Jurkat whole cell lysate (20 µg)
Lane 4: Hu brain whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab32571 observed at 125 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab32571 was shown to specifically recognize PYK2 in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when PYK2 knockout samples were examined. Wild-type and PYK2 knockout samples were subjected to SDS-PAGE. Ab32571 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/Immunofluorescence analysis of PC12 cells labelling PYK2 with unpurified ab32571 at a 1/100 dilution.
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All lanes : Anti-PYK2 antibody [YE353] (ab32571) at 1/10000 dilution (purified)
Lane 1 : Ramos cell lysate
Lane 2 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 116 kDa
Observed band size: 116 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Anti-PYK2 antibody [YE353] (ab32571) at 10000 cells (purified) + RAW264.7 cell lysate at 20 µg
Secondary
Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 116 kDa
Observed band size: 116 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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All lanes : Anti-PYK2 antibody [YE353] (ab32571) at 1/2000 dilution (purified)
Lane 1 : Mouse brain tissue lysate
Lane 2 : Rat brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 116 kDa
Observed band size: 116 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Anti-PYK2 antibody [YE353] (ab32571) at 1/5000 dilution (unpurified) + Jurkat cell lysate
Predicted band size: 116 kDa
Observed band size: 116 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebral cortex tissue labelling PYK2 with purified ab32571 at 1/300. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse cerebral cortex tissue labelling PYK2 with purified ab32571 at 1/300. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat cerebral cortex tissue labelling PYK2 with purified ab32571 at 1/300. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human brain tissue labelling PYK2 with unpurified ab32571 at a 1/250 dilution.
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Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling PYK2 with purified ab32571 at 1/60. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
Control: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (29)
ab32571 has been referenced in 29 publications.
- Lei W et al. Disulfiram-copper activates chloride currents and induces apoptosis with tyrosine kinase in prostate cancer cells. Asia Pac J Clin Oncol 18:e46-e55 (2022). PubMed: 33608991
- Zheng J et al. Pyk2 suppresses contextual fear memory in an autophosphorylation-independent manner. J Mol Cell Biol 13:808-821 (2022). PubMed: 34529077
- Chen KQ et al. Quanzhenyiqitang Reverses LPS-Induced Inflammation via Inhibiting PYK2/p38MAPK/HDAC2/CK2 Signaling Pathway in Rat Alveolar Macrophage. Evid Based Complement Alternat Med 2022:7857022 (2022). PubMed: 35047050
- Zhang C et al. High expression of PYK2 is associated with poor prognosis and cancer progression in early-stage cervical carcinoma. Medicine (Baltimore) 101:e31178 (2022). PubMed: 36253980
- Zhang Y et al. Silencing Nogo-B receptor inhibits penile corpus cavernosum vascular smooth muscle cell apoptosis of rats with diabetic erectile dysfunction by down-regulating ICAM-1. PLoS One 14:e0220715 (2019). PubMed: 31442237