Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
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Validation data
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- Product number
- PA5-65135 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PACT Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Immunogen sequence: KLAKHRAAEAA INILKANASI CFAVPDPLMP DPSKQPKNQL NPIGSLQELA IHHGWRLPEY TLSQEGGPAH KREYTTICRL ESFMETGK Highest antigen sequence identity to the following orthologs - mouse 100%, rat 100%.
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.3 mg/mL
- Storage
- Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of PACT in human cell line RT-4 and human cell line U-251 MG. Samples were probed using a PACT Polyclonal Antibody (Product # PA5-65135).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockout of PRKRA was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR762000_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of PRKRA was performed by loading 30 µg of HeLa wild type (Lane 1), HeLa Cas9 (Lane 2) andHeLa PRKRA 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-PACT Polyclonal Antibody (Product # PA5-65135, 0.3 µg/mL dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:5000 dilution) using the iBright™ FL 1500 (Product # A44115). 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 PRKRA. Uncharacterized bands were observed in all the samples between ~45-80 kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of PACT using PACT Polyclonal Antibody (Product # PA5-65135) (A) shows similar pattern to an independent PACT Polyclonal Antibody (B).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent staining of PACT in human cell line U-2 OS shows localization to cytosol. Samples were probed using a PACT Polyclonal Antibody (Product # PA5-65135).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical staining of PACT in human lateral ventricle shows strong cytoplasmic positivity in neuronal cells. Samples were probed using a PACT Polyclonal Antibody (Product # PA5-65135).