| 货号 | 7770-0104 |
| 同种亚型 | Polyclonal IgG |
| 反应种属 | Sheep |
| 来源宿主 | Rabbit |
| 应用 | E, IF, P |
| 供应商 | Bio-Rad Antibodies |
| 运输条件 | |
| 存放说明 | Store at +4oC or at -20oC if preferred. Storage in frost-free freezers is not recommended. This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use. |
| 本官网所有报价均为常温或者蓝冰运输价格，如有产品需要干冰运输，需另外加收干冰运输费。 |
Published customer image: Rabbit antiSheep prolactin antibody (7770-0104) used for the evaluation of prolactin content of secreted microvesicles by western blotting. Image caption: RHBDL4-induced stabilization of higher molecular weight forms leads to their detection in secreted microvesicles. (a) proTGFα-FLAG or CD44-FLAG were co-expressed in Hek293T cells with RHBDL4 (R4) or the catalytically inactive RHBDL4 mutant (R4-SA) in presence of 10 μM BB94. After removing cell debris, conditioned media were subjected to ultracentrifugation. The 37-kDa form of proTGFα-FLAG and a subpopulation of CD44-FLAG were recovered in the 175,000?×?g pellet fraction (p), whereas the trimmed FLAG-tagged TGFα pro-peptide (indicated by asterisk) was in the supernatant fraction (sn). t, total cell extracts. (b) proTGFα and CD44 from the 175,000?×?g pellet fraction as obtained in (a) float in a sucrose gradient indicating that they are secreted by microvesicles. Soluble secreted prolactin serves as negative control as it is found in the supernatant (sn) fraction only. Sucrose density ranges from 1.06 g/l for the top (fraction 1) to 1.25 g/l (fraction 9); sp, sucrose gradient pellet. (c) Released proTGFα-FLAG (black arrow) is trapped by a His-tag that has been introduced between TACE-cleavage site and the juxtamembrane region. In contrast, FLAG-tagged pro-peptide (asterisk) is not bound. From: Wunderle L, Knopf JD, Kühnle N, Morlé A, Hehn B, Adrain C, Strisovsky K, Freeman M, Lemberg MK. Rhomboid intramembrane protease RHBDL4 triggers ER-export and non-canonical secretion of membrane-anchored TGFα. Sci Rep. 2016 Jun 6;6:27342. | |
Published customer image: Rabbit antiSheep prolactin antibody (7770-0104) used for the evaluation of prolactin content of Hek293 cell lysates by western blotting. Image caption: RHBDL4-mediated proTGFα-secretion boost is enhanced by transactivation. (a) Exosome secretion of proTGFα-FLAG in COS7 cells ectopically expressing GRPR is stimulated by PMA or bombesin (bbs), mimicking the effect caused by RHBDL4 overexpression. In absence of BB94 (BB), transactivation by PMA and bbs also affects the extent of pro-peptide cleavage subsequently reducing the amount of secreted pro-peptide (asterisk) in the media fraction. (b) Upon transactivation of COS7 cells with bbs, the secreted 37-kDa form of proTGFα-FLAG was processed to the 6-kDa mature form via a number of intermediates. Of note, due to a low expression the ER-resident 28-kDa form of proTGFα-FLAG was not detected (cf. Fig. 1a,g). (c) Time course of TGFα secretion after PMA stimulation of Hek293T ectopically expressing proTGFα-FLAG was performed in presence of BB94 (BB, 20 μM), DCI (100 μM) or both. (d) PMA and bbs-induced proTGFα-FLAG secretion boost can be suppressed by RHBDL4 knockdown in Hek293 T-REx cells expressing an inducible shRNA but not in control cells. This effect is rescued by ectopically expressed mouse RHBDL4 (mRHBDL4-HA). Cytosolic GFP and ectopically expressed prolactin (Prl) serve as transfection and loading controls. From: Wunderle L, Knopf JD, Kühnle N, Morlé A, Hehn B, Adrain C, Strisovsky K, Freeman M, Lemberg MK. Rhomboid intramembrane protease RHBDL4 triggers ER-export and non-canonical secretion of membrane-anchored TGFα. Sci Rep. 2016 Jun 6;6:27342. |
