Gankyrin Recombinant Rabbit Monoclonal Antibody [PSH17-80]
Catalog# HA723949
Gankyrin Recombinant Rabbit Monoclonal Antibody [PSH17-80]
-
WB
-
IF-Cell
-
IHC-P
-
FC
-
IF-Tissue
-
Human
-
Mouse
-
Rat
-
Green monkey
-
unconjugated
Overview
Product Name
Gankyrin Recombinant Rabbit Monoclonal Antibody [PSH17-80]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Recombinant protein within human Gankyrin aa 1-226.
Species Reactivity
Human, Mouse, Rat, Green monkey
Validated Applications
WB, IF-Cell, IHC-P, FC, IF-Tissue
Target Molecular Weight
Predicted band size: 24 kDa
Positive Control
MCF7 cell lysate, HeLa cell lysate, K-562 cell lysate, HEK-293 cell lysate, COS-1 cell lysate, RAW264.7 cell lysate, C6 cell lysate, HeLa, RAW264.7, human skin tissue, human testis tissue.
Conjugation
unconjugated
Clone Number
PSH17-80
Reactivity Data
| WB | IF-Cell | IHC-P | FC | IF-Tissue | |
|---|---|---|---|---|---|
| Human |
|
|
|
|
|
| Mouse |
|
|
|
|
|
| Rat |
|
|
|
||
| Green Monkey |
|
Product Features
Form
Liquid
Concentration
1 mg/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
1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:5,000
-
IF-Cell
-
1:100
-
IHC-P
-
1:1,000
-
FC
-
1:1,000
-
IF-Tissue
-
1:200
Target
Function
26S proteasome non-ATPase regulatory subunit 10 or gankyrin is an enzyme that in humans is encoded by the PSMD10 gene. First isolated in 1998 by Tanaka et al.; Gankyrin is an oncoprotein that is a component of the 19S regulatory cap of the proteasome. Structurally, it contains a 33-amino acid ankyrin repeat that forms a series of alpha helices. It plays a key role in regulating the cell cycle via protein-protein interactions with the cyclin-dependent kinase CDK4. It also binds closely to the E3 ubiquitin ligase MDM2, which is a regulator of the degradation of p53 and retinoblastoma protein, both transcription factors involved in tumor suppression and found mutated in many cancers. Gankyrin also has an anti-apoptotic effect and is overexpressed in certain types of tumor cells such as hepatocellular carcinoma.
Background References
1. Lei M et al. Gankyrin inhibits ferroptosis through the p53/SLC7A11/GPX4 axis in triple-negative breast cancer cells. Sci Rep. 2023 Dec
2. Yu M et al. Gankyrin has a potential role in embryo implantation via activation of STAT3. Reproduction. 2022 Feb
Subcellular Location
Cytoplasm, Nucleus.
Synonyms
26S proteasome non-ATPase regulatory subunit 10 antibody
26S proteasome regulatory subunit p28 antibody
Ankyrin repeat protein antibody
dJ889N15.2 antibody
Gankyrin antibody
Hepatocellular carcinoma-associated protein p28 II antibody
p28 antibody
p28(GANK) antibody
Proteasome (prosome, macropain) 26S subunit, non ATPase, 10 antibody
Proteasome 26S S10 antibody
ExpandImages
-
Western blot analysis of Gankyrin on different lysates with Rabbit anti-Gankyrin antibody (HA723949) at 1/5,000 dilution.
Lane 1: MCF7 cell lysate
Lane 2: HeLa cell lysate
Lane 3: K-562 cell lysate
Lane 4: HEK-293 cell lysate
Lane 5: COS-1 cell lysate
Lane 6: RAW264.7 cell lysate
Lane 7: C6 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 24 kDa
Observed band size: 24 kDa
Exposure time: 25 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 (HA723949) at 1/5,000 dilution was used in primary antibody dilution (K1803) 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 Gankyrin with Rabbit anti-Gankyrin antibody (HA723949) at 1/100 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-Gankyrin antibody (HA723949) at 1/100 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of RAW264.7 cells labeling Gankyrin with Rabbit anti-Gankyrin antibody (HA723949) at 1/100 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-Gankyrin antibody (HA723949) at 1/100 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 (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human skin tissue with Rabbit anti-Gankyrin antibody (HA723949) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723949) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-Gankyrin antibody (HA723949) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723949) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Flow cytometric analysis of RAW264.7 cells labeling Gankyrin.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA723949, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"