Detects human Serpin A1/ alpha1‑Antitrypsin in direct ELISAs and Western blots. In Western blots, no cross‑reactivity with recombinant human Serpin A3, A4, A5, C1, F1, recombinant mouse Serpin D1, or F2 is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG_2A

Scientific Data Images for Human Serpin A1/ alpha1-Antitrypsin Antibody

Western Blot Shows Serpin A1/ alpha1-Antitrypsin Specificity Using Knockout Cell Line.

Western blot shows lysates of HepG2 human hepatocellular carcinoma parental cell line and Serpin A1/ alpha1-Antitrypsin knockout HepG2 cell line (KO). Nitrocellulose membrane was probed with Mouse Anti-Human Serpin A1/ alpha1-Antitrypsin Monoclonal Antibody (Catalog # MAB1268) followed by HRP-conjugated secondary antibody. A specific band was detected for Serpin A1/ alpha1-Antitrypsin at approximately 46.7 kDa (as indicated) in the parental HepG2 cell line, but is not detectable in knockout HepG2 cell line. Primary antibody dilution used: 1 ug/ml. The Ponceau stained transfer of the blot is shown. This experiment was conducted under reducing conditions. Image, protocol, and testing courtesy of YCharOS Inc. See ycharos.com for additional details.

Detection of Serpin A1/ alpha1-Antitrypsin by Immunoprecipitation.

HepG2 human hepatocellular carcinoma cell line lysates were prepared and immunoprecipitation was performed using 2.0 µg of Mouse Anti-Human Serpin A1/ alpha1-Antitrypsin Monoclonal Antibody (Catalog # MAB1268) pre-coupled to Dynabeads Protein G. Immunoprecipitated Serpin A1/ alpha1-Antitrypsin was detected in Western Blot with a rabbit anti-Serpin A1 used at 1/5000. The Ponceau stained transfer of the blot is shown. SM=4% starting material; UB=4% unbound fraction; IP=immunoprecipitate; HC=antibody heavy chain. Image, protocol and testing courtesy of YCharOS Inc. (ycharos.com).

Detection of Human Serpin A1/ alpha1-Antitrypsin by Western Blot.

Western blot shows human plasma and lysates of human lung tissue and human kidney tissue. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Serpin A1/a1-Antitrypsin Monoclonal Antibody (Catalog # MAB1268) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (HAF018). Specific bands were detected for Serpin A1/a1-Antitrypsin at approximately 50-60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Serpin A1/ alpha1‑Antitrypsin in HepG2 Human Cell Line.

Serpin A1/a1-Antitrypsin was detected in immersion fixed HepG2 human hepatocellular carcinoma cell line using Mouse Anti-Human Serpin A1/ a1-Antitrypsin Monoclonal Antibody (Catalog # MAB1268) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counter-stained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Human Serpin A1/ alpha1‑Antitrypsin by Simple Western^TM.

Simple Western lane view shows lysates of human plasma, loaded at 0.2 mg/mL. A specific band was detected for Serpin A1/ alpha1‑Antitrypsin at approximately 65 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human Serpin A1/ alpha1‑Antitrypsin Monoclonal Antibody (Catalog # MAB1268) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Serpin A1/ alpha1-Antitrypsin in Human Prostate.

Serpin A1/ alpha1-Antitrypsin was detected in immersion fixed paraffin-embedded sections of Human Prostate using Mouse Anti-Human Serpin A1/ alpha1-Antitrypsin Monoclonal Antibody (Catalog # MAB1268) at 5 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using VisUCyte Antigen Retrieval Reagent-Basic (Catalog # VCTS021). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in epithelial cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

View 3 more images

Applications for Human Serpin A1/ alpha1-Antitrypsin Antibody

Application

Recommended Usage

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed HepG2 human hepatocellular carcinoma cell line

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of Human Prostate

Immunoprecipitation

25 µg/mL
Sample: Conditioned cell culture medium spiked with Recombinant Human Serpin A1/ alpha1-Antitrypsin (Catalog # 1268-PI), see our available Western blot detection antibodies

Simple Western

10 µg/mL
Sample: Human plasma

Western Blot

1 µg/mL
Sample: Human plasma, human lung tissue, and human kidney tissue

Reviewed Applications

Read 5 reviews rated 4.6 using MAB1268 in the following applications:

ELISA (2 Reviews)
Immunocytochemistry/Immunofluorescence (1 Review)
Immunoprecipitation (1 Review)
Western Blot (1 Review)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Background: Serpin A1/alpha 1-Antitrypsin

Serpin A1 is the archetypal member of the Serpin superfamily of the serine protease inhibitors (1). As one of the most abundant proteinase inhibitors in the circulation, it is synthesized in the liver and secreted into the bloodstream with the major function to protect tissues against neutrophil elastase. A severe Serpin A1 deficiency leads to several clinical complications such as pulmonary emphysema, juvenile hepatitis, cirrhosis, and hepatocellular carcinoma (2). The deficiency is caused by point mutations in naturally occurring Serpin A1 variants (over 70 are known). For example, the Z variant (Glu342 to Lys) forms intracellular inclusion bodies, is not secreted, and leads to a severe Serpin A1 deficiency (3).

References

Silverman, G.A. et al. (2001) J. Biol. Chem. 276:33293.
Barbour, K.W. et al. (2002) Genomics 80:515.
Lomas, D.A. et al. (2002) Biochem. Soc. Trans. 30:89.

Alternate Names

A1AT, alpha 1-Antitrypsin, alpha 1-Proteinase Inhibitor

Entrez Gene IDs

5265 (Human)

Gene Symbol

SERPINA1

UniProt

P01009

Additional Serpin A1/alpha 1-Antitrypsin Products

Product Documents for Human Serpin A1/ alpha1-Antitrypsin Antibody

Product Datasheets

Download Product Datasheets

COA

SDS

Download SDS

Product Specific Notices for Human Serpin A1/ alpha1-Antitrypsin Antibody

For research use only

Citations
Reviews

Human Serpin A1/ alpha1-Antitrypsin Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB1268

Key Product Details

Validated by

Species Reactivity

Validated:

Cited:

Applications