GWL Recombinant Rabbit Monoclonal Antibody [PSH07-69]
Catalog# HA722884
GWL Recombinant Rabbit Monoclonal Antibody [PSH07-69]
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WB
-
IF-Cell
-
IP
-
Human
-
Green monkey
-
unconjugated
Overview
Product Name
GWL Recombinant Rabbit Monoclonal Antibody [PSH07-69]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human GWL aa 1-350.
Species Reactivity
Human, Green monkey
Validated Applications
WB, IF-Cell, IP
Target Molecular Weight
Predicted band size: 97 kDa
Positive Control
HeLa cell lysate, LNCaP cell lysate, HepG2 cell lysate, MDA-MB-231 cell lysate, COS-1 cell lysate, HeLa.
Conjugation
unconjugated
Clone Number
PSH07-69
Reactivity Data
| WB | IF-Cell | IP | |
|---|---|---|---|
| Human |
|
|
|
| Green Monkey |
|
||
| Mouse | |||
| Rat |
Product Features
Form
Liquid
Concentration
1mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:2,000
-
IF-Cell
-
1:50
-
IP
-
1-2μg/sample
Target
Function
Serine/threonine kinase that plays a key role in M phase by acting as a regulator of mitosis entry and maintenance. Acts by promoting the inactivation of protein phosphatase 2A (PP2A) during M phase: does not directly inhibit PP2A but acts by mediating phosphorylation and subsequent activation of ARPP19 and ENSA at 'Ser-62' and 'Ser-67', respectively. ARPP19 and ENSA are phosphatase inhibitors that specifically inhibit the PPP2R2D (PR55-delta) subunit of PP2A. Inactivation of PP2A during M phase is essential to keep cyclin-B1-CDK1 activity high. Following DNA damage, it is also involved in checkpoint recovery by being inhibited. Phosphorylates histone protein in vitro; however such activity is unsure in vivo. May be involved in megakaryocyte differentiation.
Background References
1. Gall-Duncan T et al. Antagonistic roles of canonical and Alternative-RPA in disease-associated tandem CAG repeat instability. Cell. 2023 Oct.
2. Yang J et al. Sequential genome-wide CRISPR-Cas9 screens identify genes regulating cell-surface expression of tetraspanins. Cell Rep. 2023 Feb.
Subcellular Location
Cytoplasm, cytoskeleton, microtubule organizing center, centrosome, Nucleus, Cleavage furrow.
UNIPROT
Synonyms
2700091H24Rik
C88295
FLJ14813
GREATWALL
Greatwall protein kinase
GW
GWL
GWL_HUMAN
hGWL
MAST-L
ExpandImages
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Western blot analysis of GWL on different lysates with Rabbit anti-GWL antibody (HA722884) at 1/2,000 dilution.
Lane 1: HeLa cell lysate
Lane 2: LNCaP cell lysate
Lane 3: HepG2 cell lysate
Lane 4: MDA-MB-231 cell lysate
Lane 5: COS-1 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 97 kDa
Observed band size: 110 kDa
Exposure time: 59 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722884) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of HeLa cells labeling GWL with Rabbit anti-GWL antibody (HA722884) at 1/50 dilution.
Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-GWL antibody (HA722884) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/50 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
GWL was immunoprecipitated from 0.2 mg HeLa cell lysate with HA722884 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using HA722884 at 1/1,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input)
Lane 2: HA722884 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of HA722884 in HeLa cell lysate
Blocking/Dilution buffer: 5% NFDM/TBST
Exposure time: 40 seconds; ECL: K1802
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"