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Western blot
ImmunocytochemistryAntibody data
- Antibody Data
- Antigen structure
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- Validations
- Immunocytochemistry [1]
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- Product number
- PA5-27421 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- RAB1A Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant protein fragment
- Description
- Recommended positive controls: U87-MG, SK-N-SH, rat brain, Rab 1A tansfected 293T cell. Predicted reactivity: Mouse (100%), Rat (100%), Xenopus laevis (98%), Dog (100%), Pig (99%), Chicken (100%), Sheep (100%), Rhesus Monkey (100%), Bovine (80%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.26 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
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- Invitrogen Antibodies (provider)
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- Experimental details
- Knockout of RAF1 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR627776_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of RAF1 was performed by loading 30 µg of HeLa wild type (Lane 1), HeLa Cas9 (Lane 2) and HeLa RAF1 KO (Lane 3) whole cell extracts. The samples were electrophoresed using NuPAGE™ Novex™ 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 c-Raf Polyclonal Antibody (Product # PA5-29333, 1:2000 dilution) and Goat anti-Rabbit IgG (Heavy Chain) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:20,000 dilution) using the iBright™ FL1500 (Product # A44115). Chemiluminescent detection was performed usingSuperSignal™ 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 RAF1.
