MA5-18071

antibody from Invitrogen Antibodies

Targeting: HDAC1 GON-10, HD1, KDAC1, RPD3L1

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Recommended by provider)

Flow cytometry (Recommended by provider)

Chromatin Immunoprecipitation (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-18071

Provider

Invitrogen Antibodies

Product name

HDAC1 Monoclonal Antibody (10E2)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This target has a predicted molecular weight of 55 kDa. Actual Western blot results show a band at 57 kDa.

Reactivity

Human, Mouse, Rat

Host

Mouse

Isotype

IgG

Antibody clone number

10E2

Vial size

100 µL

Concentration

1 mg/mL

Storage

Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of HDAC1 in Hela lysates using an HDAC1 monoclonal antibody (Product # MA5-18071) at various dilutions, followed by detection using an AP-conjugated mouse IgG whole molecule antibody at a dilution of 1:2000 on a BCIP/NBT substrate.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of HDAC1 in Hela lysates using an HDAC1 monoclonal antibody (Product # MA5-18071) at various dilutions, followed by detection using an AP-conjugated mouse IgG whole molecule antibody at a dilution of 1:2000 on a BCIP/NBT substrate.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of HDAC1 was achieved by transfecting MCF7 cells with HDAC1 specific siRNAs (Silencer® select Product # s73). Western blot analysis (Fig. a) was performed using Modified whole cell extracts (1% SDS) from the HDAC1 knockdown cells (Lane 3), non-specific scrambled siRNA transfected cells (Lane 2) and untransfected cells (Lane 1). The blot was probed with Anti-HDAC1 Monoclonal Antibody (10E2) (Product # MA5-18071, 1:1000 dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). 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 HDAC1..

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-HDAC1 Polyclonal Antibody (Product # MA5-18071) and ~55kDa band corresponding to HDAC1 was observed in MCF-7, HeLa, HEK-293, NIH/3T3 and PC-12 cells. Modified whole cell extracts (1% SDS) (30 ug lysate) of MCF-7 (Lane 1), HeLa (Lane 2), HEK-293 (Lane 3), NIH/3T3 (Lane 4) and PC-12 (Lane 5) were electrophoresed using 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 (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005)..

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of HDAC1 was performed using 70% confluent log phase MCF7 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 HDAC1 Monoclonal Antibody (10E2) (Product # MA5-18071) at 1 µg/mL in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Mouse IgG (H+L), Superclonal™ Recombinant Secondary Antibody, Alexa Fluor 488 (Product # A28175) 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 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.

Chromatin Immunoprecipitation

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Chromatin Immunoprecipitation (ChIP) assay of endogenous HDAC1 protein using Anti-HDAC1 Antibody: ChIP was performed using Anti-HDAC1 Mouse Monoclonal Antibody (Product # MA5-18071, 5 µg) on sheared chromatin from HeLa cells using the MAGnify ChIP System kit (Product # 49-2024). Normal Mouse IgG was used as a negative IP control. The purified DNA was analyzed by qPCR using primers binding to CDKN1A Intron 1 and SATA satellite repeats. Data is presented as fold enrichment of the antibody signal versus the negative control IgG using the comparative CT method.