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Learn701403
antibody from Invitrogen Antibodies
Targeting: SHH
HHG1, HLP3, HPE3, MCOPCB5, SMMCI, TPT, TPTPS
Western blotAntibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [2]
- Flow cytometry [1]
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- Product number
- 701403 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SHH Recombinant Rabbit Monoclonal Antibody (23H15L1)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human, Mouse
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- 23H15L1
- Vial size
- 100 µg
- Concentration
- 0.5 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 Sonic Hedgehog in whole extracts of HepG2 lysate using a Sonic Hedgehog recombinant rabbit monoclonal antibody (Product # 701403) at a dilution of 2 µg/mL. Samples were detected using chemiluminescence (ECL). Results show a band at ~50kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-SHH Recombinant Rabbit Monoclonal Antibody (23H15L1) (Product # 701403) and a ~50 kDa band corresponding to Sonic hedgehog protein was observed in Mouse Brain and was absent in Mouse Heart and Mouse Skeletal Muscle. Tissue extracts (30 µg lysate) of Mouse Brain (Lane 1), Mouse Heart (Lane 2) and Mouse Skeletal Muscle (Lane 3) were electrophoresed using NuPAGE™ 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 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 SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of Sonic Hedgehog was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Sonic Hedgehog (23H15L1) Recombinant Rabbit Monoclonal Antibody (Product # 701403) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Sonic Hedgehog in U2OS cells using a Sonic Hedgehog recombinant rabbit monoclonal antibody (Product # 701403) followed by detection using an Alexa Fluor 488-conjugated goat anti-rabbit secondary antibody (green) (Image A). Nuclei were stained using DAPI (Image B) and actin stained with Alexa Fluor 594 phalloidin (red) (image C). Image D is a composite image showing cytoplasmic localization of SHH.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of Sonic Hedgehog in HeLa cells using a Sonic Hedgehog recombinant rabbit monoclonal antibody (Product # 701403). Cells were fixed and permeabilized using FIX & PERM (Product # GAS004) reagent, and detection was performed using an Alexa Fluor 488 goat anti-rabbit IgG (right peak) compared to an isotype control (middle peak, black) and a control without primary antibody (left peak, blue).
