Anti-C3 antibody [EPR2988]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-C3 antibody [EPR2988] (AB181147)

Intracellular Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling C3 with purified ab181147 at 1/20 dilution (5 μg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-C3 antibody [EPR2988] (AB181147)

Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling C3 with purified ab181147 at 1/250 dilution (0.4 μg/mL). Cells were fixed in 100% Methanol and permeabilized with None. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/mL). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1/1000 (2 μg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• WB

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Western blot - Anti-C3 antibody [EPR2988] (AB181147)

The molecular weights observed are consistent with what have been described in the literature (PMID : 23619360)

All lanes:

Western blot - Anti-C3 antibody [EPR2988] (ab181147) at 1/1000 dilution

Lane 1:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Human plasma lysate at 20 µg

Lane 3:

Human serum lysate at 20 µg

Lane 4:

Human spinal cord lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 187 kDa

Observed band size: 115 kDa,34 kDa,40 kDa

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• WB

CiteAb

Western blot - Anti-C3 antibody [EPR2988] (AB181147)

C3 western blot using anti-C3 antibody [EPR2988] ab181147. Publication image and figure legend from Schäfer, N., Rasras, A., et al., 2021, Front Immunol, PubMed 34819935.

ab181147 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab181147 please see the product overview.

C3aR expression was time-dependently regulated in FHR-3 stressed ARPE-19 cells. (A)C3AR mRNA expression was decreased after 5 h and increased after 24 h FHR-3 incubation. This effect could be confirmed on protein level : (B) Western blots of ARPE-19 cell lysates showed a tendency for decreased C3aR levels (54 kDa) 5 h after FHR-3 treatment. Supplementary Figure 1M shows full Western blots. (C) Decreased C3aR protein levels were detected by immunofluorescence using a specific anti-C3aR antibody (red) after 5 h FHR-3 incubation (upper panels), whereas no differences between FHR-3 stressed and unstressed controls were observed after 24 h (lower panels). Scale bars 40 μm. w/o untreated control (dotted line). Mean with standard deviation is shown. *p < 0.05. (A) Wilcoxon matched-pairs signed rank test (n = 3); (B) Ordinary one-way ANOVA, Turkey's multiple comparisons test (n = 3).

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