Detection of Human LIN-28B by Western Blot.
Western blot shows lysates of K562 human chronic myelogenous leukemia cell line. PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human LIN-28B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7726) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for LIN-28B at approximately 22 and 35‑37 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
LIN-28B in Human Liver.
LIN-28B was detected in immersion fixed paraffin-embedded sections of human liver using Sheep Anti-Human LIN-28B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7726) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in hepatocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Lin-28B (Lineage protein 28 homolog B) is a 35-37 kDa member of the Lin28 family of proteins. Although there is an accompanying Lin-28/28A molecule, Lin-28 and Lin-28B are not alternative splice forms but represent products of separate genes. Lin-28B is found in both the cytoplasm and nucleus of fetal hematopoietic stem and progenitor cells, embryonic liver and multiple tumor types. It is considered a regulator of miRNA activity. Active miRNAs arise from transcribed RNAs that are first cleaved by nuclear RNase III, next cleaved by cytoplasmic Dicer, and finally associate with Argonaute to generate a RISC RNA silencing complex. Lin-28A participates in the cytoplasmic uridylation of miRNAs on their 3’-end, blocking Dicer processing and initiating RNA degradation. Lin-28B, by contrast, acts in the nucleus and blocks the initial RNase III cleavage step. Human Lin-28B is 250 amino acids (aa) in length (SwissProt # Q6ZN17). It contains one CSD/cold shock domain (aa 29-102) plus two CCHC-type Zn-finger RNA bindingdomains (aa 127-144 and 149-166). Phosphorylation is known to occur on Ser105. There is one reported alternate start site at Met71, and there is a potential 11 aa substitution for aa 1-4. Two isoforms of LIN28B with apparent MWs of 22 kDa and 37 kDa have been reported. Over aa 83-250, human Lin-28B shares 84% aa sequence identity with mouse Lin-28B.
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