701510

antibody from Invitrogen Antibodies

Targeting: SQSTM1 A170, OSIL, p60, p62, p62B, PDB3

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

701510

Provider

Invitrogen Antibodies

Product name

SQSTM1 Recombinant Rabbit Monoclonal Antibody (11HC14LC25)

Antibody type

Monoclonal

Antigen

Other

Description

This antibody is predicted to react with bovine, feline, non-human primate, mouse, rat, ovine and porcine based on sequence homology.

Antibody clone number

11HC14LC25

Concentration

0.5 mg/mL

References

Submitted references

The pneumococcal two-component system SirRH is linked to enhanced intracellular survival of Streptococcus pneumoniae in influenza-infected pulmonary cells.

Reinoso-Vizcaíno NM, Cian MB, Cortes PR, Olivero NB, Hernandez-Morfa M, Piñas GE, Badapanda C, Rathore A, Perez DR, Echenique J
PLoS pathogens 2020 Aug;16(8):e1008761

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Composition of the Intranuclear Inclusions of Fragile X-associated Tremor/Ataxia Syndrome.

Ma L, Herren AW, Espinal G, Randol J, McLaughlin B, Martinez-Cerdeño V, Pessah IN, Hagerman RJ, Hagerman PJ
Acta neuropathologica communications 2019 Sep 3;7(1):143

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Sequestosome-1 recombinant protein (lane 1, 200 ng) and endogenous Sequestome-1 in extracts from HeLa cells treated with 60 mM chloroquinone for 16 hours (lane 2) using a Sequestome-1 Recombinant Rabbit Polyclonal Antibody (Product # 701510) at a concentration of 1 µg/mL. A rabbit Anti-tubulin antibody was blotted as a loading control and the blots were developed using chemiluminescence (ECL) method with a Goat anti-Rabbit HRP-conjugated secondary antibody (Product # G-21234). Results show a band at ~62 kDa.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-SQSTM1 Recombinant Rabbit Monoclonal Antibody (11HC14LC25) (Product # 701510) and a ~55kDa band corresponding to SQSTM1 was observed across cell lines tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1) and Hep G2 (Lane 2) were electrophoresed using NuPAGE™ 10% Bis-Tris Protein Gel (Product # NP0301BOX). 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 (0.5 µg/mL) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of SQSTM1 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with SQSTM1 Recombinant Rabbit Monoclonal Antibody (11HC14LC25) (Product # 701510) at 1 µg/mL in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing cytoplasm, endosome-like, plasma membrane and nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.