701293

antibody from Invitrogen Antibodies

Targeting: POMC ACTH, CLIP, LPH, MSH, NPP, POC

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

701293

Provider

Invitrogen Antibodies

Product name

ACTH Recombinant Rabbit Monoclonal Antibody (9H5L7)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This antibody is predicted to react with non-human primate, human, porcine and rabbit based on sequence homology.

Antibody clone number

9H5L7

Concentration

0.5 mg/mL

References

Submitted references

Immune Checkpoint Blockade Anti-PD-L1 as a Trigger for Autoimmune Polyendocrine Syndrome.

Lanzolla G, Coppelli A, Cosottini M, Del Prato S, Marcocci C, Lupi I
Journal of the Endocrine Society 2019 Feb 1;3(2):496-503

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of ACTH in whole tissue extracts from mouse brain using an ACTH recombinant rabbit monoclonal antibody (Product # 701293) at a dilution of 1 µg/mL. Samples were detected using chemiluminescence (ECL). Results show a band at ~29kDa.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Adrenocorticotropic Hormone was performed using 70% confluent log phase U-87 MG cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Adrenocorticotropic Hormone (9H5L7) Recombinant Rabbit Monoclonal Antibody (Product # 701293) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conj µgate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometry analysis of Adrenocorticotropic Hormone was done on U-87 MG cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0.25% Triton™ X-100 for 20 minutes, and blocked with 5% BSA for 30 minutes at room temperature. Cells were labeled with Adrenocorticotropic Hormone Antibody, ABfinity™ Rabbit Monoclonal (Product # 70-1293, red histogram) or with rabbit isotype control (pink histogram) at 3-5 µg/million cells in 2.5% BSA. After incubation at room temperature for 2 hours, the cells were labeled with Alexa Fluor® 488 Goat Anti-Rabbit Secondary Antibody (Product # A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10, 000 cells were acquired and analyzed for each sample using an Attune® Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Indirect ELISA analysis of ACTH in rat brain lysate coated onto the plate using an ACTH recombinant rabbit monoclonal antibody (Product # 701293) at various dilutions. A non-linear regression analysis was performed (4 PL) and LOD and LOQ for the antibody were determined.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 2. Immunofluorescence cytosolic diffuse staining pattern produced by pituitary antibodies.