Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
- Flow cytometry [1]
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Validation data
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- Product number
- PA5-14358 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- IDH1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with bovine and rat based on sequence homology.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 400 µL
- Concentration
- 2 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 using an IDH1 polyclonal antibody (Product # PA5-14358) in HepG2 cell line and mouse liver tissue lysates (35 µg per lane).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on whole cell extracts (30 µg lysate) of Hep G2 (Lane 1), MCF7 (Lane 2), U-87 MG (Lane 3), LNCaP (Lane 4), C2C12 (Lane 5), HeLa (Lane 6), tissue extracts of Rat Liver (Lane 7) and Mouse Liver (Lane 8). The blot was probed with Anti-IDH1 Polyclonal Antibody (Product # PA5-14358, 1:2000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/ml, 1:4000 dilution). A 47 kDa band corresponding to IDH1 was observed across the cell lines and tissues tested.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockout of IDH1 was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR1014479_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of IDH1 was performed by loading 50 µg of HeLa Wild Type (Lane 1), HeLa Cas9 (Lane 2) andHeLa IDH1 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 IDH1 Polyclonal Antibody (Product # PA5-14358, 1:2,000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10,000 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 IDH1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of IDH1 was achieved by transfecting Hep G2 with IDH1 specific siRNAs (Silencer® select Product # s7120, s7121). Western blot analysis (Fig. a) was performed using whole cell extracts from the IDH1 knockdown cells (lane 3), non-specific scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with IDH1 Polyclonal Antibody (Product # PA5-14358, 1:2000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25µg/ml, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to IDH1.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of HepG2 cells using an IDH1 polyclonal antibody (Product # PA5-14358) at a dilution of 1:10-50, followed by a fluor-conjugated goat anti-rabbit secondary antibody (green). Nuclei were stained with DAPI (blue).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical analysis of formalin-fixed, paraffin-embedded human hepatocarcinoma tissue using an IDH1 polyclonal antibody (Product # PA5-14358), followed by HRP-conjugated secondary antibody and DAB staining.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of 293 cells using an IDH1 polyclonal antibody (Product # PA5-14358) (right) compared to a negative control cell (left) at a dilution of 1:10-50, followed by a FITC-conjugated goat anti-rabbit antibody