PA5-28814

antibody from Invitrogen Antibodies

Targeting: INHBB

Western blot (Data presented on provider website)

Immunohistochemistry (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-28814

Provider

Invitrogen Antibodies

Product name

INHBB Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

Recommended positive controls: A431, H1299. Predicted reactivity: Mouse (100%), Rat (100%), Zebrafish (100%), Pig (100%), Chicken (100%), Bovine (100%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human

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.

References

Submitted references

Metastatic breast cancer cells overexpress and secrete miR-218 to regulate type I collagen deposition by osteoblasts.

Liu X, Cao M, Palomares M, Wu X, Li A, Yan W, Fong MY, Chan WC, Wang SE
Breast cancer research : BCR 2018 Oct 22;20(1):127

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 3 miR-218 directly targets INHBB and YY1, and increases INHBA expression. a Relative RNA level of INHBB and INHBA normalized to 18S in MDA-231 transfected with miR-218 or control miRNA mimic at 24 h and 72 h, respectively. *** P < 0.001. b Western blot analyses of inhibin betaB and inhibin betaA in mimic-transfected MDA-231 cells at 48 h. c Relative RNA level of INHBB and INHBA normalized to 18S in MDA-231-bone transfected with anti-miR-218 or control antagomiR at 24 h and 48 h, respectively.. ** P < 0.01; *** P < 0.001. d Sequence alignment of miR-218 and its predicted targets INHBB and YY1. e psiCheck2 reporter plasmids and miR-218 mimic or its corresponding control were transfected into indicated cell lines. Renilla and firefly luciferase activity was measured at 48 h. * P < 0.05; n.s. not significant. f Relative RNA level of YY1 normalized to 18S in mimic-transfected MDA-231 cells at 24 h. *** P < 0.001. g Western blot analyses of YY1 and inhibin betaA in MDA-231 cells co-transfected with miR-218 mimic and YY1-overexpressing plasmid, or the corresponding controls at 72 h. h psiCheck2 reporter plasmids and miR-218 mimic or its corresponding control vector were transfected into MDA-231 cells. Renilla and firefly luciferase activity was measured at 48 h. ** P < 0.01; n.s. not significant

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Fig. 4 miR-218-regulated inhibin betaA affects SMAD2/3 signaling in a cell-dependent manner. a and b Western blot analyses of phospho-SMAD2/3 in MDA-231 ( a ) or MC3T3 ( b ) that was serum-starved overnight and treated with EV-depleted CM collected from indicated cells for 30 min or 2 h. CM producing cell lines were transfected, PBS washed 48 h after transfection and then incubated with serum-free medium overnight before CM collection and EV depletion by ultracentrifugation. c Western blot analyses of inhibin betaA, inhibin betaB, and inhibin alpha in CM from indicated cell lines that was concentrated with medium concentrator columns. Arrows indicate the position of antigen. Control mimic-transfected MDA-231 whole cell lysate (WCL) was used as positive control for inhibin betaA and inhibin betaB. d Western blot analyses of inhibin betaA monomer or dimer (activin A) in CM from indicated cell lines under non-reducing condition and without boiling the samples. Bottom and top arrows indicate inhibin betaA monomer and dimer, respectively. e Western blot analyses of inhibin alpha in MC3T3 CM concentrated from 20 ml CM. WCL of MCF10A was used as a positive control. f and g Western blot analyses of phospho-SMAD2/3 in MDA-231 ( f ) or MC3T3 ( g ) that was serum-starved overnight and treated with CM collected from indicated cells for 30 min or 2 h. For MDA-231 cells ( f ), recombinant inhibin alpha protein was added to CM as indicated. For MC3T3 cells ( g ), anti-inhibin alpha antibody

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Additional file 8: Figure S3. miR-218 regulated inhibin beta expression and enhanced SMAD signaling in MCF-7 cells. a Western blot analyses of inhibin betaB and inhibin betaA in miRNA mimic-transfected MCF-7 cells at 48 h after transfection. b Western blot analyses of phospho-SMAD2/3 in MCF-7 cells that were serum-starved overnight and then treated with CM collected from indicated cells for 30 min. The CM-producing cells were transfected, PBS washed at 48 h after transfection, and then incubated with serum-free medium overnight before CM collection. c Western blot analysis of inhibin alpha in MCF-7 cells. WCL of MCF10A was used as a positive control. (PDF 145 kb)