Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
- Chromatin Immunoprecipitation [1]
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Validation data
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- Product number
- MA5-14821 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- PSIP1 Monoclonal Antibody (E.493.5)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- It is not recommended to aliquot this antibody.
- Reactivity
- Human, Mouse, Rat
- Host
- Rabbit
- Isotype
- IgG
- Antibody clone number
- E.493.5
- Vial size
- 100 µL
- Concentration
- 9 µg/mL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of LEDGF in extracts from NIH/3T3 and C6 cells using LEDGF monoclonal antibody (Product # MA5-14821).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot was performed using Anti-PSIP1 Rabbit Monoclonal Antibody (Product # MA5-14821) and a 75 kDa band corresponding to PSIP1 was observed across cell lines tested and was decreased upon Retinoic acid treatment in SH-SY5Y cell line. Modified whole cell extracts (1%SDS) (30 µg lysate) of SH-SY5Y (Lane1), SH-SY5Y treated with Retinoic acid (10uM for 8 Days) (Lane 2), Jurkat (Lane 3), EB-2 (Lane 4), NTERA-2 (Lane 5), MCF-7 (Lane 6), MDA-MB-231 (Lane 7) and T-47D (Lane 8) were electrophoresed using Novex® NuPAGE® 4-12% Bis-Tris gel (Product # NP0342BOX). 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 (1:1000 dilution) 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
- Immunofluorescence analysis of PSIP was performed using SH-SY5Y and differentiated SH-SY5Y cells (Retinoic acid 10 uM, 8 Days). 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 PSIP Rabbit Monoclonal Antibody (E.493.5) (Product # MA5-14821) at 1:250 dilution in 0.1% BSA and incubated overnight at 4 degree 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 SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d and e represents the merged image showing nuclear localization. Differentiated SH-SY5Y cells show lower levels of PSIP (Panel d) as compared to undifferentiated SH-SY5Y cells (Panel e). Panel f 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 analysis of LEDGF in paraffin-embedded human breast carcinoma using a LEDGF monoclonal antibody (Product # MA5-14821) in the presence of control peptide (left) or antigen-specific peptide (right).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Enrichment of endogenous PSIP1 protein at specific gene loci using Anti-PSIP1 Antibody: Chromatin Immunoprecipitation (ChIP) was performed using Anti-PSIP1 Rabbit Monoclonal Antibody (Product # MA5-14821, 5 µg) on sheared chromatin from 2 million SHSY5Y 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 with PCR primer pairs over CCNB1, HSPB1, HOXA9 and SAT2. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.