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Recombinant Rabbit Monoclonal
Anti-HA Tag Rabbit mAb
Catalog Number: PTM-5389
$ 130

Clone Number: JRMR-10036-1

Host: Rabbit Clonality: Recombinant Monoclonal

Applications： WB ICC/IF IP ChIP

Reactivity: All

Synonyms: /

Product Size
100 μl
Quantity

Shipping: Ambient temperature

Order online or send purchase order to info@ptmbio.com

FAQ Technical Support Protocols

General Information
Isotype IgG
Conjugate Unconjugated
Synonyms /
UniProt ID

/

Immunogen Synthetic peptide corresponding to the HA tag sequence: YPYDVPDYA
MW (kDa) /
Specificity Anti-HA Tag Rabbit mAb recognizes the exogeneously expressed HA-tagged proteins.
Product Usage Information
Applications Dilution Recommended Species
WB 1:5000 - 1:10000 All
ICC/IF 1:1000 - 1:2000 All
IP 1:25 All
ChIP 6 μg per 2x106 cells All
Properties
Purity Protein A purified
Constituents PBS, Glycerol, BSA
Storage Store at -20°C. Avoid freeze/thaw cycles.
Stability Stable for 12 months from date of receipt/reconstitution.
Target Information

Background

Human influenza hemagglutinin (HA) is a surface glycoprotein required for the infectivity of the human influenza virus. The HA tag is derived from the HA-molecule corresponding to amino acids 98-106. It has been extensively used as a general epitope tag in expression vectors. Many recombinant proteins have been engineered to express the HA tag, which does not appear to interfere with the bioactivity or the biodistribution of the recombinant protein. This tag facilitates the detection, isolation, and purification of the protein of interest.

Cellular location

/

Images
WB

Lysates: 1. HEK-293T cells transfected with an empty vector (vector control), whole cell lysate; 2. HEK-293T cells transfected with a TIM4 (WT) expression vector (42 kDa) containing a HA tag, whole cell lysate
Protein loading amount: 10 μg
Blocking buffer: 5% NFDM/TBST
Primary Ab dilution: 1:2000, 1:5000, 1:10000
Primary Ab incubation: 2 hours at room temperature
Secondary Ab: Goat Anti-Rabbit IgG H&L pAb (HRP Conjugate)
Exposure time: 60 seconds
Predicted band size: 43 kDa
Observed band size: 50 kDa
ICC/IF

Samples: A) HEK-293T cells transfected with a control plasmid; B) HEK-293T cells transfected with an HA tagged plasmid
Fixative: 100% Ice-cold methanol
Permeabilization: 0.1% Triton X-100
Primary Ab dilution: 1:2000
Primary Ab incubation condition: 4°C overnight
Secondary Ab: Goat Anti-Rabbit IgG
Nuclear counter stain: DAPI (blue)
Counter stain: Tubulin (red)
Comment: The green color represents the positive signal observed with PTM-5389.
IP

IP of whole cell lysates from HEK-293T cells transfected with an HA-tagged TIM4 (WT) expression vector
Lane 1: 5% Input
Lane 2: IP with PTM-5389
Lane 3: IP with Rabbit mAb IgG Isotype Control
IP Ab incubation condition: 1:100 dilution, 4°C overnight
Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
WB primary Ab incubation condition: PTM-5389, 1:1000 dilution, 2 hours at room temperature
Secondary Ab: Anti-Rabbit IgG for IP (HRP Conjugate)
Exposure time: 120 seconds
Predicted band size: 43 kDa
Observed band size: 50 kDa
ChIP

Sample: 293F cells transfected with an HA-tagged HA-TEST expression vector (20 kDa)
Cross-linking conditions: Cells were fixed with EGS for 30 minutes, followed by formaldehyde for 10 minutes.
Amount of chromatin per IP: 2×106 cells
Amount of Ab per IP: 6 μg
Beads type and amount per IP: 50 μL of Protein A MagBeads
Description: Chromatin immunoprecipitations were performed with PTM-5389 Anti-HA Tag Rabbit mAb, PTM-7343 Anti-MAX Rabbit mAb, and Rabbit mAb IgG Isotype Control. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human DHX40 promoter, CLTC promoter, TTC16 promoter, FLYWCH2 promoter, and HES1 intron 2 regions. The data are presented as enrichment of each sample relative to the total amount of input chromatin at each amplicon.

Research Use

For research use only, not for use in diagnostic procedures.

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