Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [4]
- Immunocytochemistry [2]
- Immunohistochemistry [1]
- Chromatin Immunoprecipitation [1]
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- Product number
- PA5-52049 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- SMARCD1 Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Recombinant full-length protein
- Description
- Immunogen sequence: MGPAPGQGLY RSPMPGAAYP RPGMLPGSRM TPQGPSMGPP GYGGNPSVRP GLAQSGMDQS RKRPAPQQIQ QVQQQAVQNR NHNAKK Highest antigen sequence identity to the following orthologs: Mouse - 100%, Rat - 100%.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Concentration
- 0.15 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 SMARCD1 in Lane 1: Marker (kDa) 230, 110, 82, 49, 32, 26, 18; Lane 2: Human cell line RT-4; Lane 3: Human cell line U-251MG sp. Samples were probed using a SMARCD1 Polyclonal Antibody (Product # PA5-52049).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of SMARCD1 in Lane 1: NIH-3T3 cell lysate (Mouse embryonic fibroblast cells); Lane 2: NBT-II cell lysate (Rat Wistar bladder tumour cells). Samples were probed using a SMARCD1 Polyclonal Antibody (Product # PA5-52049).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Knockdown of SMARCD1 was achieved by transfecting HeLa with SMARCD1 specific siRNAs (Silencer® select Product # s13152, s13151). Western blot analysis (Fig. a) was performed using whole cell extracts from the SMARCD1 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with SMARCD1 Polyclonal Antibody (Product # PA5-52049, 0.4µg/ml) 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 SMARCD1.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-SMARCD1 Polyclonal Antibody (Product # PA5-52049) and a 58kDa band corresponding to SMARCD1 was observed in all the tested cell models and tissue lysates. Whole cell lysates (30ug lysate) of U-2 OS (Lane 1), A549 (Lane 2), Jurkat (Lane 3), HeLa (Lane 4), SH-SY5Y (Lane 5), and tissue extracts (30ug lysate) of Mouse Brain (Lane 6) were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris 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 (0.4µg/ml) and detected by chemiluminescence Goat Anti-Rabbit IgG Secondary Antibody, HRP conjugate (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent staining of SMARCD1 in human cell line A-431 shows positivity in vesicles & nucleus but excluded from the nucleoli. Samples were probed using a SMARCD1 Polyclonal Antibody (Product # PA5-52049).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of SMARCD1 was performed using HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with SMARCD1 Polyclonal Antibody (Product # PA5-52049) at 5 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing nuclear localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemical staining of SMARCD1 in human testis tissue shows strong nuclear positivity in cells in seminiferus ducts. Samples were probed using a SMARCD1 Polyclonal Antibody (Product # PA5-52049).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Chromatin Immunoprecipitation (ChIP) assay of endogenous SMARCD1 protein using Anti-SMARCD1 Antibody: ChIP was performed using Anti-SMARCD1 Rabbit Polyclonal Antibody (Product # PA5-52049, 5 µg) on sheared chromatin from MCF7 cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Rabbit IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to pS2 and CCND1 promoters and SAT2 satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.