Recombinant Anti-Haptoglobin antibody [EPR22856-212] (ab256454)

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Western blot - Anti-Haptoglobin antibody [EPR22856-212] (ab256454)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR22856-212] to Haptoglobin
Suitable for: Flow Cyt (Intra), WB, IHC-P, IHC-Fr
Reacts with: Mouse, Rat, Human

Carrier Free

Product name

Anti-Haptoglobin antibody [EPR22856-212]
See all Haptoglobin primary antibodies
Description

Rabbit monoclonal [EPR22856-212] to Haptoglobin
Host species

Rabbit
Specificity

This antibody is not recommended for human IHC-P.
Tested applications

Suitable for: Flow Cyt (Intra), WB, IHC-P, IHC-Frmore details
Unsuitable for: ICC/IF or IP
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Mouse serum, plasma, liver and lung lysate. Rat plasma and liver lysate. Human serum and plasma and liver lysate. IHC-P: Mouse lung, mouse liver and rat liver tissues. IHC-Fr: Mouse lung tissue; rat lung tissue. Flow Cyt (intra): Hepa1-6 cells.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR22856-212
Isotype

IgG
Research areas

Alternative Versions
- Anti-Haptoglobin antibody [EPR22856-212] - BSA and Azide free (ab261918)
Compatible Secondaries
Immunohistochemistry kits
- Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab256454 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
Flow Cyt (Intra)		1/700.
WB		1/1000. Predicted molecular weight: 45 kDa.
IHC-P		1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. This antibody is not recommended for human IHC-P.
IHC-Fr		1/500.

Notes
Flow Cyt (Intra) 1/700.
WB 1/1000. Predicted molecular weight: 45 kDa.
IHC-P 1/5000. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. This antibody is not recommended for human IHC-P.
IHC-Fr 1/500.

Application notes

Is unsuitable for ICC/IF or IP.

Function

As a result of hemolysis, hemoglobin is found to accumulate in the kidney and is secreted in the urine. Haptoglobin captures, and combines with free plasma hemoglobin to allow hepatic recycling of heme iron and to prevent kidney damage. Haptoglobin also acts as an Antimicrobial; Antioxidant, has antibacterial activity and plays a role in modulating many aspects of the acute phase response. Hemoglobin/haptoglobin complexes are rapidely cleared by the macrophage CD163 scavenger receptor expressed on the surface of liver Kupfer cells through an endocytic lysosomal degradation pathway.
Uncleaved haptoglogin, also known as zonulin, plays a role in intestinal permeability, allowing intercellular tight junction disassembly, and controlling the equilibrium between tolerance and immunity to non-self antigens.
Tissue specificity

Expressed by the liver and secreted in plasma.
Involvement in disease

Anhaptoglobinemia
Sequence similarities

Belongs to the peptidase S1 family.
Contains 1 peptidase S1 domain.
Contains 2 Sushi (CCP/SCR) domains.
Cellular localization

Secreted.
Target information above from: UniProt accession P00738 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 3240 Human
- Entrez Gene: 15439 Mouse
- Entrez Gene: 24464 Rat
- Omim: 140100 Human
- SwissProt: P00738 Human
- SwissProt: Q61646 Mouse
- SwissProt: P06866 Rat
- Unigene: 513711 Human
- Unigene: 702099 Human
- Unigene: 26730 Mouse
- Unigene: 10950 Rat
see all
Alternative names
- Binding peptide antibody
- BP antibody
- Haptoglobin alpha chain antibody
- Haptoglobin alpha(1S) beta antibody
- Haptoglobin alpha(2FS) beta antibody
- Haptoglobin beta chain antibody
- Haptoglobin, alpha polypeptide antibody
- Haptoglobin, beta polypeptide antibody
- HP antibody
- HP2 ALPHA2 antibody
- HP2ALPHA2 antibody
- HPA1S antibody
- HPT antibody
- HPT_HUMAN antibody
- MGC111141 antibody
- Zonulin antibody
see all

Western blot - Anti-Haptoglobin antibody [EPR22856-212] (ab256454)

All lanes : Anti-Haptoglobin antibody [EPR22856-212] (ab256454) at 1/1000 dilution

Lane 1 : Human serum
Lane 2 : Human plasma
Lane 3 : Human liver lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

Predicted band size: 45 kDa
Observed band size: 42,45 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID:19805376, 16543473).

Exposure time: Lanes 1-2: 3 seconds; Lane 3: 10 seconds.
Western blot - Anti-Haptoglobin antibody [EPR22856-212] (ab256454)

All lanes : Anti-Haptoglobin antibody [EPR22856-212] (ab256454) at 1/2000 dilution

Lane 1 : Mouse serum
Lane 2 : Mouse plasma
Lane 3 : Mouse liver lysate
Lane 4 : Mouse lung lysate
Lane 5 : Rat plasma
Lane 6 : Rat liver lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 45 kDa
Observed band size: 36,42,45,52 kDa why is the actual band size different from the predicted?

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature (PMID:19805376, 16543473).

Exposure time: Lanes 1-2:3 6 seconds; Lanes 3-4: 3minutes; Lane 5: 3 seconds; Lane 6: 59 seconds.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Haptoglobin antibody [EPR22856-212] (ab256454)

Immunohistochemical analysis of paraffin-embedded rat liver tissue labeling Haptoglobin with ab256454 at 1/5000 dilution (0.149 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining in rat liver. The section was incubated with ab256454 for 10 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Haptoglobin antibody [EPR22856-212] (ab256454)

Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling Haptoglobin with ab256454 at 1/5000 dilution (0.149 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining in mouse liver (PMID: 14697247). The section was incubated with ab256454 for 10 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Haptoglobin antibody [EPR22856-212] (ab256454)

Immunohistochemical analysis of paraffin-embedded mouse lung tissue labeling Haptoglobin with ab256454 at 1/5000 dilution (0.149 ug/ml) followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining in mouse lung (PMID: 14697247). The section was incubated with ab256454 for 10 mins at RT. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.
Flow Cytometry (Intracellular) - Anti-Haptoglobin antibody [EPR22856-212] (ab256454)

Intracellular flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Hepa1-6 (mouse hepatoma epithelial cell) cells labeling Haptoglobin with ab256454 at 1/700 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.
Immunohistochemistry (Frozen sections) - Anti-Haptoglobin antibody [EPR22856-212] (ab256454)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen section of rat lung tissue labeling Haptoglobin with ab256454 at 1/500 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). The nuclear counter stain is DAPI (blue). Positive staining on alveolar epithelial type II cells of rat lung (PMID: 19501987).

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Immunohistochemistry (Frozen sections) - Anti-Haptoglobin antibody [EPR22856-212] (ab256454)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen section of mouse lung tissue labeling Haptoglobin with ab256454 at 1/500 dilution, followed by ab150077 AlexaFluor^®488 Goat anti-Rabbit secondary at 1/1000 dilution (green). The nuclear counter stain is DAPI (blue). Positive staining on alveolar epithelial type II cells and bronchi of mouse lung (PMID: 19501987).

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).
Anti-Haptoglobin antibody [EPR22856-212] (ab256454)

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab256454? Please let us know so that we can cite the reference in this datasheet.

ab256454 has been referenced in 1 publication.

Li G et al. Dysregulation of ferroptosis may involve in the development of non-small-cell lung cancer in Xuanwei area. J Cell Mol Med 25:2872-2884 (2021). PubMed: 33528895

Customer reviews and Q&As

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Immunohistochemistry kits

Isotype control

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (1)

Customer reviews and Q&As