Anti-Chromogranin A antibody

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody (AB15160)

IHC image of Chromogranin A staining in a section of formalin-fixed paraffin-embedded normal human pancreas* performed on a Leica Biosystems BOND® RX instrument. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab15160, 1/500 dilution, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• WB

Unknown

Western blot - Anti-Chromogranin A antibody (AB15160)

Chromogranin A is easily fragmented and both bands observed in the Western Blot above are believed to correspond to Chromogranin A.

All lanes:

Western blot - Anti-Chromogranin A antibody (ab15160) at 1/100 dilution

All lanes:

Western blot - Recombinant Human Chromogranin A protein (His tag N-Terminus) (<a href='/en-us/products/proteins-peptides/recombinant-human-chromogranin-a-protein-ab85486'>ab85486</a>) at 0.01 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 50 kDa

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Exposure time: 30s

• IHC

CiteAb

Immunohistochemistry - Anti-Chromogranin A antibody (AB15160)

Immunohistochemistry using Anti-Chromogranin A antibody, ab15160. Publication image from Owen, R. P. et al., 2018, Nat Commun, 30323168. Legend direct from paper.

LEFTY1 and OLFM4 are mainly expressed in Barrett’s oesophagus cells that do not express differentiated secretory cell markers. a Upper panel, cluster consensus matrix of BO cells from 4 BO patients (n = 371 cells). Blue-to-red colours denote the frequency with which cells are grouped together in 250 repeat clusterings of simulated technical replicates (see Methods). Clusters (B1-B4) are indicated by the coloured bars below. Lower panel, heatmaps showing expression of selected functionally relevant genes that are differentially expressed between cell clusters (>4 fold change, FDR <1e-5). b Immunohistochemical staining of MUC2, LEFTY1 and CHGA in sections derived from the same BO resection specimen. Black arrows indicate goblet cells on all sections (positively stained for MUC2; negative for LEFTY1 and CHGA). Scale bars are 50 µm. c Immunohistochemical staining of LEFTY1 in an OSG from a normal squamous endoscopic biopsy obtained from a patient with BO. Scale bars are 300 µm and 50 µm in enlarged image

• IHC-Fr

CiteAb

Immunohistochemistry (Frozen sections) - Anti-Chromogranin A antibody (AB15160)

Chromogranin A immunohistochemistry-immunofluorescence using Anti-Chromogranin A antibody ab15160. Publication image and figure legend from Jones, B. C., Cala G., et al., 2021, Pediatr Surg Int, PubMed 33495862.

a Endoscopic biopsy of gastric mucosa from a paediatric patient prior to processing. b, c Characterisation by immunofluorescence of gastric mucosa from endoscopic biopsies. Mucin 6 (MUC6) in green, chromogranin A (CHRA) in red, Mucin 5AC (MUC5AC) in cyan, pepsinogen C (PGC) in magenta, and nuclei in blue (Hoechst). Scale bars 50 μm. d Bright field images of gastric glands isolated from endoscopic biopsies forming epithelial gastric organoids when cultured in chemically defined medium (detailed in methods). Scale bars 100 μm

• WB

CiteAb

Western blot - Anti-Chromogranin A antibody (AB15160)

Chromogranin A western blot using Anti-Chromogranin A antibody ab15160. Publication image and figure legend from Simpson, P. D., Eipper, B. A., et al., 2015, J Biol Chem, PubMed 26296884.

Regulation of CgA C-terminal immunoactivity in H727 cells is rapid and highly responsive to changes in oxygen concentration.A, upper panel, IB of extracts from cells in normoxia or at 1% O2 for indicated times. The CgA signal observed after a 15-min exposure of cells to 1% O2 is 11 ± 16% S.D. of the signal at 4 h. Quantitation based on n = 3 biological replicates. Lower panel, graph displaying the CgA IB data over the time course at 1% O2 (n = 3), indicating a linear response. B, protein synthesis is required for the CgA response to changes in oxygen concentration. Upper panel, IB of extracts from either control normoxic (N) or hypoxic cells (H, 1% O2 for 180 min) treated or not at t = 0 with 0.1 mm cycloheximide (CHX). Note that HIF-1α protein induction in hypoxia is prevented by cycloheximide treatment consistent with the known requirement for ongoing protein synthesis (39). Lower panel, hypoxic cells (3 h at 1% O2) were incubated under hypoxic conditions for a further 135 min prior to harvest with and without the addition of CHX (0.1 mm) for the indicated times. C, cells were incubated in normoxia, hypoxia (3 h at 1% O2), or hypoxia followed by re-oxygenation to 21% O2 for the indicated times. The HIF-1α IB signal is gone after 25 min reoxygenation. CgA C-terminal IB signal is also lost upon reoxygenation with 61 ± 4.6% S.D. of the t = 0 signal remaining at 125 min. Quantitation based on n = 3 biological replicates. D, IB of extracts from cells incubated for 3 h either in normoxia (21% O2) or graded hypoxia (7, 5, and 3% O2).

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• IHC-P

CiteAb

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chromogranin A antibody (AB15160)

Chromogranin A immunohistochemistry using Anti-Chromogranin A antibody ab15160. Publication image and figure legend from Oh, S. & Park, J. T. 2019, Anim Cells Syst (Seoul), PubMed 31231585.

Immunohistochemical profiles of the abnormal pancreatic region at 6 and 12 months of age. (A-C) Histological profiles in Tg (ela3I-CRE; LSL-KRASG12D) fish (A and B). Boxed areas indicate regions depicted at higher magnification in adjacent images. In the control group (w/o ela3I-CRE), normal histology was observed (C). Scale bars : 50 μm. (D-F) Enhanced chromogranin A staining was observed in the KRASG12D-induced pancreatic tumor (D and E), whereas infrequent chromogranin A staining was observed in the endocrine region of the pancreas in the control group (w/o ela3I-CRE) (F). Scale bars : 50 μm. (G-I) The chromogranin A-positive tissues were also positive for GFP staining (G and H), whereas no GFP staining was observed in the control group (w/o ela3I-CRE). Scale bars : 50 μm. (J-L) Trichrome staining to identify the regions of fibrosis and sclerosis. The blue collagen fibers (black arrows) demonstrated that the invading carcinoma triggered fibrosis/sclerosis (J and K). In the control group (w/o ela3I-CRE), typical fibrotic changes were observed in the pancreas (L). Scale bars : 50 μm. (M-R) KRASG12D-induced pancreatic tumors showed widespread labeling for both phospho-AKT (M and N) and phospho-ERK (P and Q), in contrast to infrequent AKT and ERK phosphorylation in controls (w/o ela3I-CRE) (O and R). Scale bar : 50 µm.