Human/Mouse PAK3 Antibody

Catalog #: AF6897 Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
AF6897
AF6897-SP

Detection of Human PAK3 by Western Blot.

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Human/Mouse PAK3 Antibody Summary

Species Reactivity

Human, Mouse

Specificity

Detects human PAK3 in direct ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human (rh) PAK1, rhPAK1B, rhPAK2, rhPAK4, rhPAK6, and rhPAK7 is observed.

Source

Polyclonal Sheep IgG

Purification

Antigen Affinity-purified

Immunogen

E. coli-derived recombinant human PAK3
Leu138-Gln255
Accession # O75914

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.25 µg/mL

See below

Simple Western

5 µg/mL

Human hippocampus

Immunohistochemistry

5-15 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human PAK3 by Western Blot. Western blot shows lysates of human brain (hypothalamus) tissue. PVDF membrane was probed with 0.25 µg/mL of Sheep Anti-Human PAK3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6897) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for PAK3 at approximately 65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunohistochemistry

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PAK3 in Mouse Embryo. PAK3 was detected in immersion fixed frozen sections of mouse embryo (15 d.p.c.) using Sheep Anti-Human PAK3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6897) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to the cytoplasm of neuronal cells. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

Immunohistochemistry

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PAK3 in Human Brain. PAK3 was detected in immersion fixed paraffin-embedded sections of human brain (hippocampus) using Sheep Anti-Human PAK3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6897) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to neuronal cell bodies and processes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Simple Western

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Detection of Human PAK3 by Simple Western^TM. Simple Western shows lysates of human hippocampus, loaded at 0.5 mg/ml. A specific band was detected for PAK3 at approximately 65 kDa (as indicated) using 5 µg/mL of Sheep Anti-Human/Mouse PAK3 Antigen Affinity-purified Polyclonal Antibody (Catalog # af6897). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.2 mg/mL. For liquid material, refer to CoA for concentration.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: PAK3

PAK3 (p21-activaed kinase 3; also beta -PAK, PAK3b and oligophrenin-3) is a cytoplasmic 68 kDa member of PAK group I, STE20 subfamily, STE Ser/Thr protein kinase family of molecules. It is expressed in neurons of cortical layers II, III and V, the arcuate nucleus, and neurons in the dorsal raphe plus locus coeruleus. PAK3 mediates axon and dentrite arborization, providing volume to the mass of the CNS. It binds to active GTPases which induce PAK autophosphorylation and activation. Human PAK3(b) is 559 amino acids (aa) in length. It contains one GTPase binding domain (aa 68-128) plus a protein kinase catalytic region (aa 283-534). Acetylation occurs on Lys535, and there are at least three potential phosphorylation sites. Alternate splicing generates three isoform variants. There is a 65 kDa short form that shows a deletion of aa 93-107 (PAK3a), a 69 kDa long form that shows a 21 aa substitution for aa 93-107 (PAK3c), and a 72 kDa very long isoform that contains the aforementioned 21 aa "substitution" inserted onto PAK3b after Thr92 (PAK3cb). PAK3b, c and cb are all constitutively active, and show only modest binding affinity towards GTPases. Over aa 138-255, human PAK3(b) shares 94% aa identity with mouse PAK3(b).

Long Name

p21-activated Kinase 3

Entrez Gene IDs

5063 (Human); 18481 (Mouse)

Alternate Names

beta-PAK; bPAK; CDKN1A; EC 2.7.11; EC 2.7.11.1; hPAK3; mental retardation, X-linked 47; MRX30; MRX47; oligophrenin-3; OPHN3; p21 (CDKN1A)-activated kinase 3; p21 protein (Cdc42/Rac)-activated kinase 3; p21-activated kinase 3; PAK3; PAK-3; PAK3beta; serine/threonine-protein kinase PAK 3