PA5-29584

antibody from Invitrogen Antibodies

Targeting: A2M CPAMD5, FWP007, S863-7

Western blot (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-29584

Provider

Invitrogen Antibodies

Product name

A2M Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: 293T, A431, Jurkat, human plasma.

Concentration

1 mg/mL

References

Submitted references

Insight into the human pathodegradome of the V8 protease from Staphylococcus aureus.

Frey AM, Chaput D, Shaw LN
Cell reports 2021 Apr 6;35(1):108930

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot analysis of A2M was performed by separating 30 µg of human tissue extract by 5% SDS-PAGE. Proteins were transferred to a membrane and probed with a A2M Polyclonal Antibody (Product # PA5-29584) at a dilution of 1:10000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using A2M Polyclonal Antibody (Product # PA5-29584). Sample (30 µg of whole cell lysate). Lane A: A431 . 5% SDS PAGE. A2M Polyclonal Antibody (Product # PA5-29584) diluted at 1:1,000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-A2M Polyclonal Antibody (Product # PA5-29584) and a 180 kDa band corresponding to Alpha-2-macroglobulin was observed across tested samples. Whole cell extracts (Fig. a) (40 µg lysate) of Hep G2 (Lane 1), Hep G2 treated with Protein transport inhibitor (1X for 4 hours) (Lane 2), SK-BR-3 (Lane 3), SK-BR-3 treated with Protein transport inhibitor (1X for 4 hours) (Lane 4), Reh (Lane 5), PC-3 (Lane 6), Human Plasma (Fig. b) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580).

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 4. Validation of V8 serum substrates identified by N-terminomics Serum was treated with varying concentrations of V8. Linear schematics depict the primary structure of proteins, with domains from UniProt/UniRef indicated. Cleavage sites inferred by N-terminomics are indicated by red markers, the scissile bond/V8 cleavage site is indicated by an asterisk with surrounding residues shown. Fragments and their ~predicted mass (calculated using ProtParam), with degradation products visible in corresponding western blots, are shown on schematics for serotransferrin, alpha-2-macroglobulin, SERPINA1, and SERPINC1. Prothrombin produced no visible degradation products; instead, the intact form and native products (produced by proteolytic activation) are depicted and visible in the untreated serum blot. AP1, activation peptide fragment 1; AP2, activation peptide fragment 2; light, thrombin light chain; heavy, thrombin heavy chain; TF1, Transferrin-like domain 1; TF2, transferrin-like domain 2; Ig, Ig-like fold; BR, bait region; RBD, receptor binding domain; RCL, reactive center loop. See also Figure S4 .