PA5-75591

antibody from Invitrogen Antibodies

Targeting: MOB2 HCCA2

Provider product page for PA5-75591

Western blot

Other assay

Antibody data

Antibody Data
Antigen structure
References [1]
Comments [0]
Validations
Other assay [7]

Submit

Product number: PA5-75591 - Provider product page
Provider: Invitrogen Antibodies
Product name: MOB2 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Synthetic peptide
Description: The antibody was affinity-purified from rabbit antiserum by affinity-chromatography using epitope-specific immunogen and the purity is > 95% (by SDS-PAGE).
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 1 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

MOB2 protein structure - PA5-75591 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 1 MOB2 expression is decreased in glioblastoma and associated with adverse prognosis. a Normal brain ( n = 8), glioblastoma (GBM, n = 19) and low grade glioma ( n = 16) samples were detected by Immunohistochemistry (IHC). Scale bar = 100 mum. Data are presented as mean +- SEM (*** p < 0.001). b The mRNA levels of MOB2 were analyzed in glioblastoma and low grades gliomas according to the TCGA datasets. Significance level was determined using the non-parametric Mann-Whitney test. Data are presented as mean +- SEM (*** p < 0.001). c MOB2 was analyzed in glioblastoma and normal brain samples according to GSE4290 and GSE16011 microarray datasets. Significance level was determined using the Non-parametric Mann-Whitney test. Data are presented as mean +- SEM (*** p < 0.001). d MOB2 protein expression in normal brain and glioma cells was detected by IB. e Kaplan-Meier survival curves for correlation between MOB2 mRNA expression and survival of gliomas patients in the TCGA RNA-seq dataset.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 2 MOB2 is a tumor suppressor in GBM. a IB analysis of the efficiency of stable knockdown of MOB2 in LN-229 and T98G cells (shCON and shMOB2)or stable overexpression of MOB2 in SF-539 and SF-767 cells (pCDH-VEC and pCDH-MOB2), GAPDH was used as a loading control. b Cell proliferation rate of MOB2-depleted LN-229 and T98G cells was measured by BrdUrd labeling. c Representative images and quantification of migration and invasion of MOB2-depleted LN-229 and T98G cells. Scale bar = 200 mum. d Cell proliferation of MOB2-depleted LN-229 and T98G cells was measured by colony formation assays. e Cell proliferation rate of SF-539 and SF-767 cells stably expressing MOB2 was measured by BrdUrd labeling. f Representative images and quantification of migration and invasion of MOB2-overexpressed SF-539 and SF-767 cells. Scale bar = 200 mum. g Cell proliferation of SF-539 and SF-767 cells stably expressing MOB2 was measured by colony formation assays. h , i Chick embryos with CAM tumors formed from MOB2-depleted LN-229 and T98G cells ( h ) or MOB2-overexpressed SF-539 and SF-767 cells ( i ) were dissected and the size of CAM tumors was calculated. A minimum of ten embryos per condition was analyzed. j Enlarged tumor volumes in MOB2-overexpressed SF-767 cells vs control mice, were measured weekly (7 weeks). All experiments were performed as three independent experiments. Data are presented as mean +- SEM (* p < 0.05, ** p < 0.01, *** p < 0.001).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 4 FAK-Akt signaling pathway is activated by knocking down MOB2. a , b IB with specific antibodies against p-FAK, FAK, p-Akt, Akt, p-Paxillin, Paxillin, and MOB2 in MOB2-depleted LN-229 and T98G cells ( a ), and SF-539 and SF-767 cells stably overexpressing MOB2 ( b ). GAPDH was used as a loading control. c LN-229 and T98G cells stably knockdowning of MOB2 or control cells were treated with vehicle (DMSO) or PF566271, IB analysis for p-FAK, FAK, p-Akt, Akt, and MOB2. GAPDH was used as a loading control. d LN-229 and T98G cells stably knockdowning of MOB2 or control cells were treated with vehicle (DMSO) or MK2206, IB analysis for p-FAK, FAK, p-Akt, Akt, and MOB2. GAPDH was used as a loading control. e LN-229 and T98G cells stably knockdowning of MOB2 or control cells were treated the same as in c , the ability of migration and invasion was measured by Transwell chamber assay. f LN-229 and T98G cells stably knockdowning of MOB2 or control cells were treated the same as in d , the ability of migration and invasion was measured by Transwell chamber assay. g SF-539 and SF-767 cells with stable overexpression of MOB2 or control cells were treated with vehicle (DMSO) or SC79, the ability of migration and invasion was measured by Transwell chamber. h LN-229 and T98G cells were transfected with two siFAK (siFAK-1 and siFAK-2) or siControl (siCON) oligonucleotides for 48 h, then IB analysis for FAK, GAPDH was used as a loading control. i LN-229 and T98G cells with a stable knockdo

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 5 Integrin plays an important role in MOB2-regulated FAK/Akt signaling. a LN-229 and T98G cells with a stable knockdown of MOB2 or control cells were treated with vehicle (DMSO) or Cyclo-RGDfK (C-RGD) IB analysis for p-FAK, FAK, p-Akt, Akt, and MOB2, GAPDH was used as a loading control. b LN-229 and T98G cells with a stable knockdown of MOB2 were treated with vehicle (DMSO), AZD9291 (2 muM), BAY-11-7086 (10 muM), C118-9 (20 muM), C-RGD (1 muM), FSK (10 muM), LY364947 (10 muM), MK2206 (2 muM), PD98059 (10 muM), PF566271 (2 muM) or Rapamycin (2 muM), IB analysis for p-FAK, FAK, p-Akt, and Akt, GAPDH was used as a loading control. c - e LN-229 and T98G cells with a stable knockdown of MOB2 or control cells were treated with vehicle (DMSO) or AZD9291 ( c ), C-RGD ( d ) and LY364947 ( e ), the ability of migration and invasion was measured by Transwell chamber assay. All experiments were performed as three independent experiments. Data were presented as mean +- SEM (*** p < 0.001).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 6 MOB2 is critical for cAMP/PKA-mediated inhibition of cell migration and invasion in glioma cells. a LN-229 and T98G cells with a stable knockdown of MOB2 or control cells were treated with vehicle (DMSO) or Forskolin (FSK), IB analysis for p-FAK, FAK, p-Akt, Akt and MOB2, GAPDH was used as a loading control. b SF-539 and SF-767 cells were treated with vehicle (DMSO) or Forskolin (FSK), IB analysis for p-FAK, FAK, p-Akt, Akt, p-CREB and CREB, GAPDH was used as a loading control. c SF-539 and SF-767 cells with stable overexpression of MOB2 or control cells were treated with vehicle (DMSO) or H89, IB analysis for p-FAK, FAK, p-Akt, and Akt, GAPDH was used as a loading control. d LN-229 and T98G cells with a stable knockdown of MOB2 or control cells were treated with vehicle (DMSO) or Forskolin (FSK), and then the ability of migration and invasion was measured by Transwell chamber assay. e SF-539 and SF-767 cells with stable overexpression of MOB2 cells or control cells were treated with vehicle (DMSO) or H89, and then the ability of migration and invasion was measured by Transwell chamber assay. All experiments were performed as three independent experiments. Data are presented as mean +- SEM (* p < 0.05, ** p < 0.01, *** p < 0.001).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 7 MOB2 binds to PKA and enhances PKA activity. a LN-229 and T98G cells with a stable knockdown of MOB2 or control cells were treated with vehicle (DMSO) or Forskolin (FSK), IB analysis for p-CREB, CREB and MOB2, GAPDH was used as a loading control. b SF-539 and SF-767 cells with stable overexpression of MOB2 or control cells were treated with vehicle (DMSO) or SQ22536, IB analysis for p-CREB, CREB and MOB2, GAPDH was used as a loading control. c LN-229 and T98G whole-cell lysates (WCL) collected from 10 cm 2 dishes were subjected to immunoprecipitation (IP) with an anti-MOB2 antibody or an IgG control. d , e 293T cells transfected with GFP-tagged PKA (GFP-PKA) and V5-tagged MOB2 (V5-MOB2). Cell lysates were subjected to immunoprecipitation (IP) with anti-GFP ( d ) or anti-V5 antibody ( e ) and immunoblotted (IB) with the indicated antibodies. f Hs683, LN-18 and U87MG cells were treated with vehicle (DMSO) or Forskolin (FSK), IB analysis for MOB1 and MOB2, GAPDH was used as a loading control. g LN-229 and T98G cells were treated with vehicle (DMSO) or H89, IB analysis for MOB1 and MOB2, GAPDH was used as a loading control. All experiments in this figure were performed as three independent experiments.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Supplementary Figure 4. The effects of MOB2 depletion on the FAK/Akt signaling pathway were rescued by either wild type (WT) MOB2 or the MOB2-H157A mutant