MA1-26774

antibody from Invitrogen Antibodies

Targeting: CTSL CTSL1, FLJ31037

ELISA

Immunohistochemistry

Antibody data

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Validation data

Product number: MA1-26774 - Provider product page
Provider: Invitrogen Antibodies
Product name: Cathepsin L Monoclonal Antibody (33/2)
Antibody type: Monoclonal
Antigen: Purifed from natural sources
Description: Recommended positive controls: mink lung cell.
Reactivity: Human, Mouse, Rat
Host: Mouse
Isotype: IgG
Antibody clone number: 33/2
Vial size: 100 µL
Concentration: 4.3 mg/mL
Storage: Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

CTSL protein structure - MA1-26774 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot of Cathepsin L in whole cell extract of Mv 1 Lu cultured mink lung using a Cathepsin L monoclonal antibody (Product # MA1-26774) at a dilution of 1:400 (Lane A) compared to a negative control with only secondary antibody (Lane B). The antibody was developed with a Goat anti-mouse IgG (Fab), AlkPhos Conjugate and NBT/BCIP substrate.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockout of Cathepsin L was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR1042354_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of Cathepsin L was performed by loading 50 µg of A549 wild type (Lane 1), A549 Cas9 (Lane 2) andA549 Cathepsin L KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-Cathepsin L Monoclonal Antibody (33/2) (Product # MA1-26774, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:10000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to Cathepsin L.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of Cathepsin L was achieved by transfecting A549 with Cathepsin L specific siRNAs (Silencer® select Product # S223364, S3754). Western blot analysis (Fig. a) was performed using Whole cell extracts from the Cathepsin L knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with Cathepsin L Monoclonal Antibody (33/2) (Product # MA1-26774, 1:1000 dilution ) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of all three expected bands of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to Cathepsin L.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-Cathepsin L Monoclonal Antibody (33/2)(Product # MA1-26774) and a 25kDa band corresponding to the mature form of Cathepsin L was observed across cell lines and tissue extracts tested. An additional faint bands around 41kDa which corresponds to the Procathepsin L and 31kDa which also corresponds to the mature form of Cathepsin L was observed in A549. Whole cell extracts (30 µg lysate) of A549 (Lane 1), Hep G2 (Lane 2), MCF 10A (Lane 3) and tissue extracts of Mouse Kidney (Lane 4) and Mouse Liver (Lane 5) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).