Anti-HMGB2 antibody [EPR6302] (ab133540)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6302] to HMGB2
- Suitable for: WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Overview
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Product name
Anti-HMGB2 antibody [EPR6302]
See all HMGB2 primary antibodies -
Description
Rabbit monoclonal [EPR6302] to HMGB2 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species IHC-P HumanWB Human -
Immunogen
Synthetic peptide corresponding to residues in the Human HMGB2 protein (UniProt ID: P26583).
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Positive control
- WB: HEK-293T, HAP1, K562, HL-60, C6, RAW 264.7, and PC-12 cell lysates. IHC-P: Human breast tissue and Human colon carcinoma tissue.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C. -
Dissociation constant (KD)
KD = 2.97 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.2
Preservative: 0.05% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Tissue culture supernatant -
Clonality
Monoclonal -
Clone number
EPR6302 -
Isotype
IgG -
Research areas
Images
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All lanes : Anti-HMGB2 antibody [EPR6302] (ab133540) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : HMGB2 knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 24 kDa
Observed band size: 24 kDaLanes 1- 2: Merged signal (red and green). Green - ab133540 observed at 24 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab133540 was shown to react with HMGB2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266358 (knockout cell lysate ab257156) was used. Wild-type HEK-293T and HMGB2 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133540 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry analysis of paraffin embedded Human breast tissue labelling HMGB2 with ab133540 at 1/250 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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All lanes : Anti-HMGB2 antibody [EPR6302] (ab133540) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : HMGB2 knockout HAP1 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 24 kDa
Observed band size: 24 kDaLanes 1 - 4: Merged signal (red and green). Green - ab133540 observed at 24 kDa. Red - loading control, ab9484, observed at 37 kDa.
ab133540 was shown to recognize HMGB2 in wild-type HAP1 cells as signal was lost at the expected MW in HMGB2 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and HMGB2 knockout samples were subjected to SDS-PAGE. Ab133540 and ab9484 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
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Immunohistochemistry analysis of paraffin embedded Human colon carcinoma tissue labelling HMGB2 with ab133540 at 1/250 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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All lanes : Anti-HMGB2 antibody [EPR6302] (ab133540) at 1/1000 dilution
Lane 1 : K562 cell lysate
Lane 2 : HL-60 cell lysate
Lane 3 : C6 cell lysate
Lane 4 : RAW 264.7 cell lysate
Lane 5 : PC-12 cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 24 kDa
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