Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (ab255277)

Rabbit Recombinant Monoclonal HMGA1 antibody. Carrier free. Suitable for IP, ChIP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse samples.

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Images

ChIP - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (AB255277), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	ChIP	WB	ICC/IF	IHC-Fr	Flow Cyt (Intra)	IHC-P
Human	Tested	Expected	Tested	Tested	Not recommended	Tested	Tested
Mouse	Tested	Tested	Expected	Expected	Not recommended	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Mouse	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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8 products for Alternative Version

2 products for Alternative Product

Target data

See full target information HMGA1

Function

HMG-I/Y bind preferentially to the minor groove of A+T rich regions in double-stranded DNA. It is suggested that these proteins could function in nucleosome phasing and in the 3'-end processing of mRNA transcripts. They are also involved in the transcription regulation of genes containing, or in close proximity to A+T-rich regions.

Alternative names

HMGIY, HMGA1, High mobility group protein HMG-I/HMG-Y, HMG-I(Y), High mobility group AT-hook protein 1, High mobility group protein R, High mobility group protein A1

Recommended products

Rabbit Recombinant Monoclonal HMGA1 antibody. Carrier free. Suitable for IP, ChIP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Human, Mouse samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR22421-101
Purification technique: Affinity purification Protein A
Specificity: The Mouse recommendation is based on IP and ChIP results. We do not guarantee WB and IHC for Mouse.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab255277 is the carrier-free version of Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930.

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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 batch production

For more information, read more on recombinant antibodies.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

10 product images

ChIP - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (ab255277)
Chromatin was prepared from EL4 cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min.
The ChIP was performed with 25 μg of chromatin, 5 μg of Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930 (red) or 5 μg of rabbit normal IgG (gray) was added to 20 μl of Protein A/G sepharose beads. The immunoprecipitated DNA was quantified by real time PCR (sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930).
Immunoprecipitation - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (ab255277)
HMGA1 was immunoprecipitated from 0.35 mg HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate with Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used at 1/5000 dilution.
Lane 1: HCT 116 whole cell lysate 10 μg (Input).
Lane 2: Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930 IP in HCT 116 whole cell lysate.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930 in HCT 116 whole cell lysate.
Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: 40 seconds.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930).
All lanes: Immunoprecipitation - Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade (Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930)
Predicted band size: 12 kDa
Observed band size: 17 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (ab255277)
Immunohistochemical analysis of paraffin-embedded human testis tissue labeling HMGA1 with Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in germ cells of human testis (PMID:17935122) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930).
Immunocytochemistry/ Immunofluorescence - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (ab255277)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NCCIT (human pluripotent embryonic carcinoma epithelial cell) cells labeling HMGA1 with Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in NCCIT cell line. Low expression cell line: MCF7?PMID: 12640109). The nuclear counter stain is DAPI (blue). Counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).
The negative control is the secondary antibody only.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930).
Flow Cytometry (Intracellular) - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (ab255277)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized MCF7 (Human breast adenocarcinoma epithelial cell, Left) / NCCIT (Human pluripotent embryonic carcinoma epithelial cell, Right) cell lines labeling HMGA1 with Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930 at 1/600 (red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), at 1/2000 dilution was used as the secondary antibody. Weak expression control: MCF7 (PMID:12640109).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (ab255277)
Immunohistochemical analysis of paraffin-embedded human pancreas cancer tissue labeling HMGA1 with Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human pancreas cancer (PMID: 18473350) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (ab255277)
Immunohistochemical analysis of paraffin-embedded human lung cancer tissue labeling HMGA1 with Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930 at 1/5000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Positive staining in human lung cancer (PMID:25344216) is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930).
Western blot - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (ab255277)
Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930).
Lanes 1 - 3: Western blot - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (ab255277) at 1/1000 dilution
Lanes 1 - 3: Western blot - Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade (Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930) at 1/1000 dilution
Lane 1: MDA-MB-231 (Human breast adenocarcinoma epithelial cell) nuclear fraction lysate at 15 µg
Lane 2: MDA-MB-231 (Human breast adenocarcinoma epithelial cell) cytoplasm fraction lysate at 15 µg
Lane 3: MDA-MB-231 (Human breast adenocarcinoma epithelial cell) membrane fraction lysate at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 12 kDa
Observed band size: 17 kDa
Exposure time: 10s
Western blot - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (ab255277)
Blocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
We recommend using 1% SDS Hot lysis prepare method to get desired Western Blot results.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930).
Lanes 1 - 4: Western blot - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (ab255277) at 1/1000 dilution
Lanes 1 - 4: Western blot - Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade (Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930) at 1/1000 dilution
Lane 1: HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method at 20 µg
Lane 2: HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysate prepared in 1% SDS Hot lysis method at 20 µg
Lane 3: MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate prepared in RIPA lysis method at 20 µg
Lane 4: MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysate prepared in 1% SDS Hot lysis method at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 12 kDa
Observed band size: 17 kDa
Exposure time: 40s
Immunoprecipitation - Anti-HMGA1 antibody [EPR22421-101] - BSA and Azide free (ab255277)
HMGA1 was immunoprecipitated from 0.35mg of EL4 (mouse lymphoma T lymphocyte) whole cell lysate with Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930 at 1/30 dilution.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930).
All lanes: Immunoprecipitation - Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade (Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930) at 1/1000 dilution
Lane 1: EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 10 µg
Lane 2: Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930 IP in EL4 (mouse lymphoma T lymphocyte) whole cell lysate at 10 µg
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-HMGA1 antibody [EPR22421-101] - ChIP Grade ab252930 in EL4 (human epidermoid carcinoma) whole cell lysate at 10 µg
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Predicted band size: 17 kDa
Observed band size: 12 kDa
Exposure time: 180s