Anti-Insulin Receptor beta antibody [C18C4]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Insulin Receptor beta antibody [C18C4] (AB69508)

IHC image of ab69508 staining in human normal kidney formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab69508, 1μg/ml, 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.

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.

This image was generated using the ascites version of the product.

• Flow Cyt

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Flow Cytometry - Anti-Insulin Receptor beta antibody [C18C4] (AB69508)

Overlay histogram showing HepG2 cells stained with ab69508 (red line). The cells were fixed with 80% methanol (5 min) and incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab69508, 1μg/1x10⁶ cells) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.

Please note that Abcam do not have any data for use of this antibody on non-fixed cells. We welcome any customer feedback.

This image was generated using the ascites version of the product.

• WB

Supplier Data

Western blot - Anti-Insulin Receptor beta antibody [C18C4] (AB69508)

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-Insulin Receptor beta antibody [C18C4] (ab69508) at 1/1000 dilution

Lane 2:

HeLa lysate (Human epithelial cell line from cervix adenocarcinoma)

Lane 3:

MCF-7 lysate (Human breast adenocarcinoma cell line)

Lane 4:

PC-12 lysate (Rat adrenal gland pheochromocytoma cell line)

Lane 5:

3T3-L1 lysate (Mouse embryonic fibroblast cell line)

Predicted band size: 95 kDa

Observed band size: 98 kDa

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• WB

CiteAb

Western blot - Anti-Insulin Receptor beta antibody [C18C4] (AB69508)

Insulin Receptor beta western blot using anti-Insulin Receptor beta antibody [C18C4] ab69508. Publication image and figure legend from Shih, C. K., Chen, C. M., et al., 2020, Food Nutr Res, PubMed 32425738.

ab69508 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 ab69508 please see the product overview.

Expression of insulin-signaling-related proteins in the muscle. Protein expression of p-IR & IR (panel a), p-Akt & Akt (panel b), and M-GLUT4 & T-GLU4 (panel c) were analyzed with Western blot of the homogenates in the gastrocnemius muscle. Data are expressed as means ± standard deviations (n = 5). p-IR, phospho-insulin receptor; IR, insulin receptor; p-Akt, phospho-protein kinase B; Akt, protein kinase B; M-GLU4, membrane glucose transporter 4; T-GLU4, total glucose transporter 4; Asterisks indicate significant difference compared to DM group. Statistical evaluation was performed using one-way ANOVA, followed by Duncan's multiple range test, p < 0.05.

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