PA5-89181

antibody from Invitrogen Antibodies

Targeting: SMARCE1 BAF57

Immunoprecipitation

Antibody data

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Validation data

Product number: PA5-89181 - Provider product page
Provider: Invitrogen Antibodies
Product name: BAF57 Polyclonal Antibody
Antibody type: Polyclonal
Antigen: Recombinant full-length protein
Description: Immunogen sequence: MSKRPSYAPP PTPAPATQMP STPGFVGYNP YSHLAYNNYR LGGNPGTNSR VTASSGITIP KPPKPPDKPL MPYMRYSRKV WDQVKASNPD LKLWEIGKII GGMWRDLTDE EKQEYLNEYE AEKIEYNESM KAYHNSPAYL AYINAKSRAE AALEEESRQR QSRMEKGEPY MSIQPAEDPD DYDDGFSMKH TATARFQRNH RLISEILSES VVPDVRSVVT TARMQVLKRQ VQSLMVHQRK LEAELLQIEE RHQEKKRKFL ESTDSFNNEL KRLCGLKVEV DMEKIAAEIA QAEEQARKRQ EEREKEAAEQ AERSQSSIVP EEEQAANKGE EKKDDENIPM ETEETHLEET TESQQNGEEG TSTPEDKESG QEGVDSMAEE GTSDSNTGSE SNSATVEEPP TDPIPEDEKK E; Positive Samples: U-251MG, MCF-7; Cellular Location: Nucleus
Reactivity: Human, Mouse, Rat
Host: Rabbit
Isotype: IgG
Vial size: 100 µL
Concentration: 0.19 mg/mL
Storage: -20° C, Avoid Freeze/Thaw Cycles

SMARCE1 protein structure - PA5-89181 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of extracts of various cell lines, using SMARCE1 Polyclonal antibody (Product # PA5-89181) at 1:1000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Exposure time: 90s.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of extracts from normal (control) and SMARCE1 knockout (KO) HeLa cells, using SMARCE1 Polyclonal antibody (Product # PA5-89181) at 1:1000 dilution. Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution. Lysates/proteins: 25ug per lane. Blocking buffer: 3% nonfat dry milk in TBST. Exposure time: 15s.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of BAF57 in extracts from normal (control) and SMARCE1 knockout (KO) HeLa cells using BAF57 Polyclonal Antibody (Product # PA5-89181) at a dilution of 1:1000. A HRP Goat Anti-Rabbit IgG (H+L) secondary antibody was used at a dilution of 1:10,000. Lysates/proteins: 25 µg per lane. Blocking buffer: 3% nonfat dry milk in TBST.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot analysis of BAF57 in extracts of various cell lines using BAF57 Polyclonal Antibody (Product # PA5-89181) at a dilution of 1:1000. A HRP Goat Anti-Rabbit IgG (H+L) secondary antibody was used at a dilution of 1:10,000. Lysates/proteins: 25 µg per lane. Blocking buffer: 3% nonfat dry milk in TBST.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Knockdown of SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily E member 1 was achieved by transfecting HeLa with SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily E member 1 specific siRNAs (Silencer® select Product # S13160, S13162 ). Additional uncharacterized bands (*) were also observed. Western blot analysis (Fig. a) was performed using Nuclear enriched extracts from the SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily E member 1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2), and untransfected cells (lane 1). The blot was probed with BAF57 Polyclonal Antibody (Product # PA5-89181, 1:1000 ) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000). 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 SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily E member 1.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot was performed using Anti-BAF57 Polyclonal Antibody (Product # PA5-89181) and a 57kDa band corresponding to SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily E member 1 was observed across cell lines and tissue tested. Additional uncharacterized bands (*) were observed across the panel. Nuclear enriched extracts (30 µg lysate) of HeLa (Lane 1), A-431 (Lane 2), NIH/3T3 (Lane 3), MCF7 (Lane 4), Raji (Lane 5), and Mouse Thymus (Lane 6) were electrophoresed using NuPAGE™ 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 the primary antibody (1:1000 dilution) 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 Novex® ECL Reagent Kit (Product # WP20005).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of HeLa cells using SMARCE1 Polyclonal antibody (Product # PA5-89181).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of BAF57 was performed in HeLa cells using BAF57 Polyclonal Antibody (Product # PA5-89181).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunocytochemistry-Immunofluorescence analysis of BAF57 was performed in HeLa cells using BAF57 Polyclonal Antibody (Product # PA5-89181).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescence analysis of BAF57 was performed using 70% confluent log phase A-431 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 BAF57 Polyclonal Antibody (Product # PA5-89181) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then with Goat anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32731) (1:2000) for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with Hoechst 33342 (Product # H1399). 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 40X magnification in CellInsight CX7 LZR High-Content Screening (HCS) Platform (Product # CX7C1115LZR).